72 results on '"Meents A"'
Search Results
2. Sample Preparation and Data Collection for High-Speed Fixed-Target Serial Femtosecond Crystallography
- Author
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Meents, Alke, Meents, Alke, Roedig, Philip, Ginn, Helen M, Pakendorf, Tim, Sutton, Geoff, Harlos, Karl, Walter, Thomas S, Meyer, Jan, Fischer, Pontus, Duman, Ramona, Vartiainen, Ismo, Reime, Bernd, Warmer, Martin, Brewster, Aaron S, Young, Iris D, Clark, Tara Michels, Sauter, Nicholas K, Sikorsky, Marcin, Nelson, Silke, Damiani, Daniel S, Alonso-Mori, Roberto, Ren, Jingshan, Fry, Elizabeth E, David, Christian, Stuart, David I, Wagner, Armin H, Meents, Alke, Meents, Alke, Roedig, Philip, Ginn, Helen M, Pakendorf, Tim, Sutton, Geoff, Harlos, Karl, Walter, Thomas S, Meyer, Jan, Fischer, Pontus, Duman, Ramona, Vartiainen, Ismo, Reime, Bernd, Warmer, Martin, Brewster, Aaron S, Young, Iris D, Clark, Tara Michels, Sauter, Nicholas K, Sikorsky, Marcin, Nelson, Silke, Damiani, Daniel S, Alonso-Mori, Roberto, Ren, Jingshan, Fry, Elizabeth E, David, Christian, Stuart, David I, and Wagner, Armin H more...
- Published
- 2022
Catalog
3. Calpeptin is a potent cathepsin inhibitor and drug candidate for SARS-CoV-2 infections
- Author
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Reinke, Patrick Y. A., de Souza, Edmarcia Elisa, Günther, Sebastian, Falke, Sven, Lieske, Julia, Ewert, Wiebke, Loboda, Jure, Herrmann, Alexander, Rahmani Mashhour, Aida, Karničar, Katarina, Usenik, Aleksandra, Lindič, Nataša, Sekirnik, Andreja, Botosso, Viviane Fongaro, Santelli, Gláucia Maria Machado, Kapronezai, Josana, de Araújo, Marcelo Valdemir, Silva-Pereira, Taiana Tainá, Filho, Antônio Francisco de Souza, Tavares, Mariana Silva, Flórez-Álvarez, Lizdany, de Oliveira, Danielle Bruna Leal, Durigon, Edison Luiz, Giaretta, Paula Roberta, Heinemann, Marcos Bryan, Hauser, Maurice, Seychell, Brandon, Böhler, Hendrik, Rut, Wioletta, Drag, Marcin, Beck, Tobias, Cox, Russell, Chapman, Henry N., Betzel, Christian, Brehm, Wolfgang, Hinrichs, Winfried, Ebert, Gregor, Latham, Sharissa L., Guimarães, Ana Marcia de Sá, Turk, Dusan, Wrenger, Carsten, Meents, Alke, Reinke, Patrick Y. A., de Souza, Edmarcia Elisa, Günther, Sebastian, Falke, Sven, Lieske, Julia, Ewert, Wiebke, Loboda, Jure, Herrmann, Alexander, Rahmani Mashhour, Aida, Karničar, Katarina, Usenik, Aleksandra, Lindič, Nataša, Sekirnik, Andreja, Botosso, Viviane Fongaro, Santelli, Gláucia Maria Machado, Kapronezai, Josana, de Araújo, Marcelo Valdemir, Silva-Pereira, Taiana Tainá, Filho, Antônio Francisco de Souza, Tavares, Mariana Silva, Flórez-Álvarez, Lizdany, de Oliveira, Danielle Bruna Leal, Durigon, Edison Luiz, Giaretta, Paula Roberta, Heinemann, Marcos Bryan, Hauser, Maurice, Seychell, Brandon, Böhler, Hendrik, Rut, Wioletta, Drag, Marcin, Beck, Tobias, Cox, Russell, Chapman, Henry N., Betzel, Christian, Brehm, Wolfgang, Hinrichs, Winfried, Ebert, Gregor, Latham, Sharissa L., Guimarães, Ana Marcia de Sá, Turk, Dusan, Wrenger, Carsten, and Meents, Alke more...
- Abstract
Several drug screening campaigns identified Calpeptin as a drug candidate against SARS-CoV-2. Initially reported to target the viral main protease (Mpro), its moderate activity in Mpro inhibition assays hints at a second target. Indeed, we show that Calpeptin is an extremely potent cysteine cathepsin inhibitor, a finding additionally supported by X-ray crystallography. Cell infection assays proved Calpeptin’s efficacy against SARS-CoV-2. Treatment of SARS-CoV-2-infected Golden Syrian hamsters with sulfonated Calpeptin at a dose of 1 mg/kg body weight reduces the viral load in the trachea. Despite a higher risk of side effects, an intrinsic advantage in targeting host proteins is their mutational stability in contrast to highly mutable viral targets. Here we show that the inhibition of cathepsins, a protein family of the host organism, by calpeptin is a promising approach for the treatment of SARS-CoV-2 and potentially other viral infections. more...
- Published
- 2023
4. Hard X-ray stereographic microscopy for single-shot differential phase imaging
- Author
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Bellucci, Valerio, Zdora, Marie Christine, Mikeš, Ladislav, Birnšteinová, Šarlota, Oberta, Peter, Romagnoni, Marco, Mazzolari, Andrea, Villanueva-Perez, Pablo, Mokso, Rajmund, David, Christian, Makita, Mikako, Cipiccia, Silvia, Uličný, Jozef, Meents, Alke, Mancuso, Adrian P., Chapman, Henry N., Vagovič, Patrik, Bellucci, Valerio, Zdora, Marie Christine, Mikeš, Ladislav, Birnšteinová, Šarlota, Oberta, Peter, Romagnoni, Marco, Mazzolari, Andrea, Villanueva-Perez, Pablo, Mokso, Rajmund, David, Christian, Makita, Mikako, Cipiccia, Silvia, Uličný, Jozef, Meents, Alke, Mancuso, Adrian P., Chapman, Henry N., and Vagovič, Patrik more...
- Abstract
The characterisation of fast phenomena at the microscopic scale is required for the understanding of catastrophic responses of materials to loads and shocks, the processing of materials by optical or mechanical means, the processes involved in many key technologies such as additive manufacturing and microfluidics, and the mixing of fuels in combustion. Such processes are usually stochastic in nature and occur within the opaque interior volumes of materials or samples, with complex dynamics that evolve in all three dimensions at speeds exceeding many meters per second. There is therefore a need for the ability to record three-dimensional X-ray movies of irreversible processes with resolutions of micrometers and frame rates of microseconds. Here we demonstrate a method to achieve this by recording a stereo phase-contrast image pair in a single exposure. The two images are combined computationally to reconstruct a 3D model of the object. The method is extendable to more than two simultaneous views. When combined with megahertz pulse trains of X-ray free-electron lasers (XFELs) it will be possible to create movies able to resolve 3D trajectories with velocities of kilometers per second. more...
- Published
- 2023
5. A polysaccharide utilization locus from the gut bacterium Dysgonomonas mossii encodes functionally distinct carbohydrate esterases
- Author
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Kmezik, Cathleen, Mazurkewich, Scott, Meents, Tomke, McKee, Lauren S., Idström, Alexander, Armeni, Marina, Savolainen, Otto, Brandén, Gisela, Larsbrink, Johan, Kmezik, Cathleen, Mazurkewich, Scott, Meents, Tomke, McKee, Lauren S., Idström, Alexander, Armeni, Marina, Savolainen, Otto, Brandén, Gisela, and Larsbrink, Johan more...
- Abstract
The gut microbiota plays a central role in human health by enzymatically degrading dietary fiber and concomitantly excreting short chain fatty acids that are associated with manifold health benefits. The polysaccharide xylan is abundant in dietary fiber but noncarbohydrate decorations hinder efficient cleavage by glycoside hydrolases (GHs) and need to be addressed by carbohydrate esterases (CEs). Enzymes from carbohydrate esterase families 1 and 6 (CE1 and 6) perform key roles in xylan degradation by removing feruloyl and acetate decorations, yet little is known about these enzyme families especially with regard to their diversity in activity. Bacteroidetes bacteria are dominant members of the microbiota and often encode their carbohydrate-active enzymes in multigene polysaccharide utilization loci (PULs). Here we present the characterization of three CEs found in a PUL encoded by the gut Bacteroidete Dysgonomonas mossii. We demonstrate that the CEs are functionally distinct, with one highly efficient CE6 acetyl esterase and two CE1 enzymes with feruloyl esterase activities. One multidomain CE1 enzyme contains two CE1 domains: an N-terminal domain feruloyl esterase, and a C-terminal domain with minimal activity on model substrates. We present the structure of the C-terminal CE1 domain with the carbohydrate-binding module that bridges the two CE1 domains, as well as a complex of the same protein fragment with methyl ferulate. The investment of D. mossii in producing multiple CEs suggests that improved accessibility of xylan for GHs and cleavage of covalent polysaccharide-polysaccharide and lignin-polysaccharide bonds are important enzyme activities in the gut environment., QC 20210914 more...
- Published
- 2021
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6. X-ray screening identifies active site and allosteric inhibitors of SARS-CoV-2 main protease
- Author
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Günther, Sebastian, Reinke, Patrick Y. A., Fernández-García, Yaiza, Lieske, Julia, Lane, Thomas J., Ginn, Helen M., Koua, Faisal H. M., Ehrt, Christiane, Ewert, Wiebke, Oberthuer, Dominik, Yefanov, Oleksandr, Meier, Susanne, Lorenzen, Kristina, Krichel, Boris, Kopicki, Janine-Denise, Gelisio, Luca, Brehm, Wolfgang, Dunkel, Ilona, Seychell, Brandon, Gieseler, Henry, Norton-Baker, Brenna, Escudero-Pérez, Beatriz, Domaracky, Martin, Saouane, Sofiane, Tolstikova, Alexandra, White, Thomas A., Hänle, Anna, Groessler, Michael, Fleckenstein, Holger, Trost, Fabian, Galchenkova, Marina, Gevorkov, Yaroslav, Li, Chufeng, Awel, Salah, Peck, Ariana, Barthelmess, Miriam, Schlünzen, Frank, Lourdu Xavier, P., Werner, Nadine, Andaleeb, Hina, Ullah, Najeeb, Falke, Sven, Srinivasan, Vasundara, França, Bruno Alves, Schwinzer, Martin, Brognaro, Hévila, Rogers, Cromarte, Melo, Diogo, Zaitseva-Doyle, Joanna J., Knoska, Juraj, Peña-Murillo, Gisel E., Mashhour, Aida Rahmani, Hennicke, Vincent, Fischer, Pontus, Hakanpää, Johanna, Meyer, Jan, Gribbon, Philip, Ellinger, Bernhard, Kuzikov, Maria, Wolf, Markus, Beccari, Andrea R., Bourenkov, Gleb, von Stetten, David, Pompidor, Guillaume, Bento, Isabel, Panneerselvam, Saravanan, Karpics, Ivars, Schneider, Thomas R., Garcia-Alai, Maria Marta, Niebling, Stephan, Günther, Christian, Schmidt, Christina, Schubert, Robin, Han, Huijong, Boger, Juliane, Monteiro, Diana C. F., Zhang, Linlin, Sun, Xinyuanyuan, Pletzer-Zelgert, Jonathan, Wollenhaupt, Jan, Feiler, Christian G., Weiss, Manfred S., Schulz, Eike-Christian, Mehrabi, Pedram, Karničar, Katarina, Usenik, Aleksandra, Loboda, Jure, Tidow, Henning, Chari, Ashwin, Hilgenfeld, Rolf, Uetrecht, Charlotte, Cox, Russell, Zaliani, Andrea, Beck, Tobias, Rarey, Matthias, Günther, Stephan, Turk, Dusan, Hinrichs, Winfried, Chapman, Henry N., Pearson, Arwen R., Betzel, Christian, Meents, Alke, Günther, Sebastian, Reinke, Patrick Y. A., Fernández-García, Yaiza, Lieske, Julia, Lane, Thomas J., Ginn, Helen M., Koua, Faisal H. M., Ehrt, Christiane, Ewert, Wiebke, Oberthuer, Dominik, Yefanov, Oleksandr, Meier, Susanne, Lorenzen, Kristina, Krichel, Boris, Kopicki, Janine-Denise, Gelisio, Luca, Brehm, Wolfgang, Dunkel, Ilona, Seychell, Brandon, Gieseler, Henry, Norton-Baker, Brenna, Escudero-Pérez, Beatriz, Domaracky, Martin, Saouane, Sofiane, Tolstikova, Alexandra, White, Thomas A., Hänle, Anna, Groessler, Michael, Fleckenstein, Holger, Trost, Fabian, Galchenkova, Marina, Gevorkov, Yaroslav, Li, Chufeng, Awel, Salah, Peck, Ariana, Barthelmess, Miriam, Schlünzen, Frank, Lourdu Xavier, P., Werner, Nadine, Andaleeb, Hina, Ullah, Najeeb, Falke, Sven, Srinivasan, Vasundara, França, Bruno Alves, Schwinzer, Martin, Brognaro, Hévila, Rogers, Cromarte, Melo, Diogo, Zaitseva-Doyle, Joanna J., Knoska, Juraj, Peña-Murillo, Gisel E., Mashhour, Aida Rahmani, Hennicke, Vincent, Fischer, Pontus, Hakanpää, Johanna, Meyer, Jan, Gribbon, Philip, Ellinger, Bernhard, Kuzikov, Maria, Wolf, Markus, Beccari, Andrea R., Bourenkov, Gleb, von Stetten, David, Pompidor, Guillaume, Bento, Isabel, Panneerselvam, Saravanan, Karpics, Ivars, Schneider, Thomas R., Garcia-Alai, Maria Marta, Niebling, Stephan, Günther, Christian, Schmidt, Christina, Schubert, Robin, Han, Huijong, Boger, Juliane, Monteiro, Diana C. F., Zhang, Linlin, Sun, Xinyuanyuan, Pletzer-Zelgert, Jonathan, Wollenhaupt, Jan, Feiler, Christian G., Weiss, Manfred S., Schulz, Eike-Christian, Mehrabi, Pedram, Karničar, Katarina, Usenik, Aleksandra, Loboda, Jure, Tidow, Henning, Chari, Ashwin, Hilgenfeld, Rolf, Uetrecht, Charlotte, Cox, Russell, Zaliani, Andrea, Beck, Tobias, Rarey, Matthias, Günther, Stephan, Turk, Dusan, Hinrichs, Winfried, Chapman, Henry N., Pearson, Arwen R., Betzel, Christian, and Meents, Alke more...
- Abstract
The coronavirus disease (COVID-19) caused by SARS-CoV-2 is creating tremendous human suffering. To date, no effective drug is available to directly treat the disease. In a search for a drug against COVID-19, we have performed a high-throughput X-ray crystallographic screen of two repurposing drug libraries against the SARS-CoV-2 main protease (Mpro), which is essential for viral replication. In contrast to commonly applied X-ray fragment screening experiments with molecules of low complexity, our screen tested alreadyapproved drugs and drugs in clinical trials. From the three-dimensional protein structures, we identified 37 compounds that bind to Mpro. In subsequent cell-based viral reduction assays, one peptidomimetic and six nonpeptidic compounds showed antiviral activity at nontoxic concentrations. We identified two allosteric binding sites representing attractive targets for drug development against SARS-CoV-2. more...
- Published
- 2021
7. X-ray screening identifies active site and allosteric inhibitors of SARS-CoV-2 main protease
- Author
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Günther, Sebastian, Reinke, Patrick, Fernández-Garciá, Yaiza, Lieske, Julia, Lane, Thomas, Ginn, Helen Mary, Koua, Faisal H. M., Ehrt, Christiane, Ewert, Wiebke, Oberthür, Dominik, Yefanov, Oleksandr, Meier, Susanne, Lorenzen, Kristina, Krichel, Boris, Kopicki, Janine-Denise, Gelisio, Luca, Brehm, Wolfgang, Dunkel, Ilona, Seychell, Brandon, Gieseler, Henry, Norton-Baker, Brenna, Escudero-Pérez, Beatriz, Domaracky, Martin, Saouane, Sofiane, Tolstikova, Aleksandra, White, Thomas A., Hänle, Anna, Groessler, Michael, Fleckenstein, Holger, Trost, Fabian, Galchenkova, Marina, Gevorkov, Yaroslav, Li, Chufeng, Awel, Salah, Peck, Ariana, Barthelmess, Miriam, Schlünzen, Frank, Xavier, Paulrajpillai Lourdu, Werner, Nadine, Andaleeb, Hina, Ullah, Najeeb, Falke, Sven, Srinivasan, Vasundara, Francą, Bruno Alves, Schwinzer, Martin, Brognaro, Hévila, Rogers, Cromarte, Melo, Diogo, Zaitseva-Doyle, Joanna J., Knoška, Juraj, Peña Murillo, Gisel E., Mashhour, Aida Rahmani, Hennicke, Vincent, Fischer, Pontus, Hakanpää, Johanna, Meyer, Jan H., Gribbon, Philip, Ellinger, Bernhard, Kuzikov, Maria, Wolf, Markus, Beccari, Aandrea Rosario, Bourenkov, Gleb, Stetten, David von, Pompidor, Guillaume, Bento, Isabel, Panneerselvam, Saravanan, Karpics, Ivars, Schneider, Thomas, Garcia-Alai, Maria Marta, Niebling, Stephan, Günther, Christian, Schmidt, Christina, Schubert, Robin, Han, Huijong, Boger, Juliane, Monteiro, Diana C.F., Zhang, Linlin, Sun, Xinyuanyuan, Pletzer-Zelgert, Jonathan, Wollenhaupt, Jan, Feiler, Christian, Weiss, Manfred, Schulz, Eike C., Mehrabi, Pedram, Karnǐcar, Katarina, Usenik, Aleksandra, Loboda, Jure, Tidow, Henning, Chari, Ashwin, Hilgenfeld, Rolf, Uetrecht, Charlotte, Cox, Russell, Zaliani, Andrea, Beck, Tobias, Rarey, Matthias, Günther, Stephan, Turk, Dusan, Hinrichs, Winfried, Chapman, Henry N., Pearson, Arwen, Betzel, Christian, Meents, Alke, Günther, Sebastian, Reinke, Patrick, Fernández-Garciá, Yaiza, Lieske, Julia, Lane, Thomas, Ginn, Helen Mary, Koua, Faisal H. M., Ehrt, Christiane, Ewert, Wiebke, Oberthür, Dominik, Yefanov, Oleksandr, Meier, Susanne, Lorenzen, Kristina, Krichel, Boris, Kopicki, Janine-Denise, Gelisio, Luca, Brehm, Wolfgang, Dunkel, Ilona, Seychell, Brandon, Gieseler, Henry, Norton-Baker, Brenna, Escudero-Pérez, Beatriz, Domaracky, Martin, Saouane, Sofiane, Tolstikova, Aleksandra, White, Thomas A., Hänle, Anna, Groessler, Michael, Fleckenstein, Holger, Trost, Fabian, Galchenkova, Marina, Gevorkov, Yaroslav, Li, Chufeng, Awel, Salah, Peck, Ariana, Barthelmess, Miriam, Schlünzen, Frank, Xavier, Paulrajpillai Lourdu, Werner, Nadine, Andaleeb, Hina, Ullah, Najeeb, Falke, Sven, Srinivasan, Vasundara, Francą, Bruno Alves, Schwinzer, Martin, Brognaro, Hévila, Rogers, Cromarte, Melo, Diogo, Zaitseva-Doyle, Joanna J., Knoška, Juraj, Peña Murillo, Gisel E., Mashhour, Aida Rahmani, Hennicke, Vincent, Fischer, Pontus, Hakanpää, Johanna, Meyer, Jan H., Gribbon, Philip, Ellinger, Bernhard, Kuzikov, Maria, Wolf, Markus, Beccari, Aandrea Rosario, Bourenkov, Gleb, Stetten, David von, Pompidor, Guillaume, Bento, Isabel, Panneerselvam, Saravanan, Karpics, Ivars, Schneider, Thomas, Garcia-Alai, Maria Marta, Niebling, Stephan, Günther, Christian, Schmidt, Christina, Schubert, Robin, Han, Huijong, Boger, Juliane, Monteiro, Diana C.F., Zhang, Linlin, Sun, Xinyuanyuan, Pletzer-Zelgert, Jonathan, Wollenhaupt, Jan, Feiler, Christian, Weiss, Manfred, Schulz, Eike C., Mehrabi, Pedram, Karnǐcar, Katarina, Usenik, Aleksandra, Loboda, Jure, Tidow, Henning, Chari, Ashwin, Hilgenfeld, Rolf, Uetrecht, Charlotte, Cox, Russell, Zaliani, Andrea, Beck, Tobias, Rarey, Matthias, Günther, Stephan, Turk, Dusan, Hinrichs, Winfried, Chapman, Henry N., Pearson, Arwen, Betzel, Christian, and Meents, Alke more...
- Abstract
The coronavirus disease (COVID-19) caused by SARS-CoV-2 is creating tremendous human suffering. To date, no effective drug is available to directly treat the disease. In a search for a drug against COVID-19, we have performed a high-throughput X-ray crystallographic screen of two repurposing drug libraries against the SARS-CoV-2 main protease (Mpro), which is essential for viral replication. In contrast to commonly applied X-ray fragment screening experiments with molecules of low complexity, our screen tested alreadyapproved drugs and drugs in clinical trials. From the three-dimensional protein structures, we identified 37 compounds that bind to Mpro. In subsequent cell-based viral reduction assays, one peptidomimetic and six nonpeptidic compounds showed antiviral activity at nontoxic concentrations. We identified two allosteric binding sites representing attractive targets for drug development against SARS-CoV-2., Deutsche Forschungsgemeinschaft (DFG), Bundesministerium für Bildung und Forschung (BMBF) more...
- Published
- 2021
8. A fixed-target platform for serial femtosecond crystallography in a hydrated environment.
- Author
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Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Seuring, C, Segelke, BW, He, W, Evans, AC, Pakendorf, T, Fischer, P, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Coleman, MA, Kuhl, TL, Frank, M, Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Seuring, C, Segelke, BW, He, W, Evans, AC, Pakendorf, T, Fischer, P, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Coleman, MA, Kuhl, TL, and Frank, M more...
- Abstract
For serial femtosecond crystallography at X-ray free-electron lasers, which entails collection of single-pulse diffraction patterns from a constantly refreshed supply of microcrystalline sample, delivery of the sample into the X-ray beam path while maintaining low background remains a technical challenge for some experiments, especially where this methodology is applied to relatively low-ordered samples or those difficult to purify and crystallize in large quantities. This work demonstrates a scheme to encapsulate biological samples using polymer thin films and graphene to maintain sample hydration in vacuum conditions. The encapsulated sample is delivered into the X-ray beam on fixed targets for rapid scanning using the Roadrunner fixed-target system towards a long-term goal of low-background measurements on weakly diffracting samples. As a proof of principle, we used microcrystals of the 24 kDa rapid encystment protein (REP24) to provide a benchmark for polymer/graphene sandwich performance. The REP24 microcrystal unit cell obtained from our sandwiched in-vacuum sample was consistent with previously established unit-cell parameters and with those measured by us without encapsulation in humidified helium, indicating that the platform is robust against evaporative losses. While significant scattering from water was observed because of the sample-deposition method, the polymer/graphene sandwich itself was shown to contribute minimally to background scattering. more...
- Published
- 2020
9. Crystallization of ApoA1 and ApoE4 nanolipoprotein particles and initial XFEL-based structural studies.
- Author
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Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Bauer, WJ, Segelke, B, He, W, Evans, AC, Crespo, N, Fischer, P, Pakendorf, T, Hennicke, V, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Kuhl, TL, Frank, M, Coleman, MA, Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Bauer, WJ, Segelke, B, He, W, Evans, AC, Crespo, N, Fischer, P, Pakendorf, T, Hennicke, V, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Kuhl, TL, Frank, M, and Coleman, MA more...
- Abstract
Nanolipoprotein particles (NLPs), also called "nanodiscs", are discoidal particles with a patch of lipid bilayer corralled by apolipoproteins. NLPs have long been of interest due to both their utility as membrane-model systems into which membrane proteins can be inserted and solubilized and their physiological role in lipid and cholesterol transport via HDL and LDL maturation, which are important for human health. Serial femtosecond crystallography (SFX) at X-ray free electron lasers (XFELs) is a powerful approach for structural biology of membrane proteins, which are traditionally difficult to crystallize as large single crystals capable of producing high-quality diffraction suitable for structure determination. To facilitate understanding of the specific role of two apolipoprotein/lipid complexes, ApoA1 and ApoE4, in lipid binding and HDL/LDL particle maturation dynamics and develop new SFX methods involving NLP membrane protein encapsulation, we have prepared and crystallized homogeneous populations of ApoA1 and ApoE4 NLPs. Crystallization of empty NLPs yields semi-ordered objects that appear crystalline and give highly anisotropic and diffuse X-ray diffraction, similar in characteristics to fiber diffraction. Several unit cell parameters were approximately determined for both NLPs from these measurements. Thus, low-background, sample conservative methods of delivery are critical. Here we implemented a fixed target sample delivery scheme utilizing the Roadrunner fast-scanning system and ultra-thin polymer/graphene support films, providing a low-volume, low-background approach to membrane protein SFX. This study represents initial steps in obtaining structural information for ApoA1 and ApoE4 NLPs and developing this system as a supporting scaffold for future structural studies of membrane proteins crystalized in a native lipid environment. more...
- Published
- 2020
10. Ptychographic X-ray speckle tracking with multi-layer Laue lens systems
- Author
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Morgan, AJ, Murray, KT, Prasciolu, M, Fleckenstein, H, Yefanov, O, Villanueva-Perez, P, Mariani, V, Domaracky, M, Kuhn, M, Aplin, S, Mohacsi, I, Messerschmidt, M, Stachnik, K, Du, Y, Burkhart, A, Meents, A, Nazaretski, E, Yan, H, Huang, X, Chu, YS, Chapman, HN, Bajt, S, Morgan, AJ, Murray, KT, Prasciolu, M, Fleckenstein, H, Yefanov, O, Villanueva-Perez, P, Mariani, V, Domaracky, M, Kuhn, M, Aplin, S, Mohacsi, I, Messerschmidt, M, Stachnik, K, Du, Y, Burkhart, A, Meents, A, Nazaretski, E, Yan, H, Huang, X, Chu, YS, Chapman, HN, and Bajt, S more...
- Abstract
The ever-increasing brightness of synchrotron radiation sources demands improved X-ray optics to utilize their capability for imaging and probing biological cells, nano-devices and functional matter on the nanometre scale with chemical sensitivity. Hard X-rays are ideal for high-resolution imaging and spectroscopic applications owing to their short wavelength, high penetrating power and chemical sensitivity. The penetrating power that makes X-rays useful for imaging also makes focusing them technologically challenging. Recent developments in layer deposition techniques have enabled the fabrication of a series of highly focusing X-ray lenses, known as wedged multi-layer Laue lenses. Improvements to the lens design and fabrication technique demand an accurate, robust, in situ and at-wavelength characterization method. To this end, a modified form of the speckle tracking wavefront metrology method has been developed. The ptychographic X-ray speckle tracking method is capable of operating with highly divergent wavefields. A useful by-product of this method is that it also provides high-resolution and aberration-free projection images of extended specimens. Three separate experiments using this method are reported, where the ray path angles have been resolved to within 4 nrad with an imaging resolution of 45 nm (full period). This method does not require a high degree of coherence, making it suitable for laboratory-based X-ray sources. Likewise, it is robust to errors in the registered sample positions, making it suitable for X-ray free-electron laser facilities, where beam-pointing fluctuations can be problematic for wavefront metrology. more...
- Published
- 2020
11. A fixed-target platform for serial femtosecond crystallography in a hydrated environment.
- Author
-
Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Seuring, C, Segelke, BW, He, W, Evans, AC, Pakendorf, T, Fischer, P, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Coleman, MA, Kuhl, TL, Frank, M, Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Seuring, C, Segelke, BW, He, W, Evans, AC, Pakendorf, T, Fischer, P, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Coleman, MA, Kuhl, TL, and Frank, M more...
- Abstract
For serial femtosecond crystallography at X-ray free-electron lasers, which entails collection of single-pulse diffraction patterns from a constantly refreshed supply of microcrystalline sample, delivery of the sample into the X-ray beam path while maintaining low background remains a technical challenge for some experiments, especially where this methodology is applied to relatively low-ordered samples or those difficult to purify and crystallize in large quantities. This work demonstrates a scheme to encapsulate biological samples using polymer thin films and graphene to maintain sample hydration in vacuum conditions. The encapsulated sample is delivered into the X-ray beam on fixed targets for rapid scanning using the Roadrunner fixed-target system towards a long-term goal of low-background measurements on weakly diffracting samples. As a proof of principle, we used microcrystals of the 24 kDa rapid encystment protein (REP24) to provide a benchmark for polymer/graphene sandwich performance. The REP24 microcrystal unit cell obtained from our sandwiched in-vacuum sample was consistent with previously established unit-cell parameters and with those measured by us without encapsulation in humidified helium, indicating that the platform is robust against evaporative losses. While significant scattering from water was observed because of the sample-deposition method, the polymer/graphene sandwich itself was shown to contribute minimally to background scattering. more...
- Published
- 2020
12. Crystallization of ApoA1 and ApoE4 nanolipoprotein particles and initial XFEL-based structural studies.
- Author
-
Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Bauer, WJ, Segelke, B, He, W, Evans, AC, Crespo, N, Fischer, P, Pakendorf, T, Hennicke, V, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Kuhl, TL, Frank, M, Coleman, MA, Shelby, ML, Shelby, ML, Gilbile, D, Grant, TD, Bauer, WJ, Segelke, B, He, W, Evans, AC, Crespo, N, Fischer, P, Pakendorf, T, Hennicke, V, Hunter, MS, Batyuk, A, Barthelmess, M, Meents, A, Kuhl, TL, Frank, M, and Coleman, MA more...
- Abstract
Nanolipoprotein particles (NLPs), also called "nanodiscs", are discoidal particles with a patch of lipid bilayer corralled by apolipoproteins. NLPs have long been of interest due to both their utility as membrane-model systems into which membrane proteins can be inserted and solubilized and their physiological role in lipid and cholesterol transport via HDL and LDL maturation, which are important for human health. Serial femtosecond crystallography (SFX) at X-ray free electron lasers (XFELs) is a powerful approach for structural biology of membrane proteins, which are traditionally difficult to crystallize as large single crystals capable of producing high-quality diffraction suitable for structure determination. To facilitate understanding of the specific role of two apolipoprotein/lipid complexes, ApoA1 and ApoE4, in lipid binding and HDL/LDL particle maturation dynamics and develop new SFX methods involving NLP membrane protein encapsulation, we have prepared and crystallized homogeneous populations of ApoA1 and ApoE4 NLPs. Crystallization of empty NLPs yields semi-ordered objects that appear crystalline and give highly anisotropic and diffuse X-ray diffraction, similar in characteristics to fiber diffraction. Several unit cell parameters were approximately determined for both NLPs from these measurements. Thus, low-background, sample conservative methods of delivery are critical. Here we implemented a fixed target sample delivery scheme utilizing the Roadrunner fast-scanning system and ultra-thin polymer/graphene support films, providing a low-volume, low-background approach to membrane protein SFX. This study represents initial steps in obtaining structural information for ApoA1 and ApoE4 NLPs and developing this system as a supporting scaffold for future structural studies of membrane proteins crystalized in a native lipid environment. more...
- Published
- 2020
13. pinkIndexer – a universal indexer for pink-beam X-ray and electron diffraction snapshots
- Author
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Gevorkov, Yaroslav, Barty, Anton, Brehm, Wolfgang, White, Thomas A., Tolstikova, Aleksandra, Wiedorn, Max Oliver, Meents, Alke, Grigat, Rolf-Rainer, Chapman, Henry N., Yefanov, Oleksandr, Gevorkov, Yaroslav, Barty, Anton, Brehm, Wolfgang, White, Thomas A., Tolstikova, Aleksandra, Wiedorn, Max Oliver, Meents, Alke, Grigat, Rolf-Rainer, Chapman, Henry N., and Yefanov, Oleksandr more...
- Abstract
A crystallographic indexing algorithm,pinkIndexer, is presented for the analysisof snapshot diffraction patterns. It can be used in a variety of contexts includingmeasurements made with a monochromatic radiation source, a polychromaticsource or with radiation of very short wavelength. As such, the algorithm isparticularly suited to automated data processing for two emerging measurementtechniques for macromolecular structure determination: serial pink-beam X-raycrystallography and serial electron crystallography, which until now lackedreliable programs for analyzing many individual diffraction patterns fromcrystals of uncorrelated orientation. The algorithm requires approximateknowledge of the unit-cell parameters of the crystal, but not the wavelengthsassociated with each Bragg spot. The use ofpinkIndexeris demonstrated byobtaining 1005 lattices from a published pink-beam serial crystallography dataset that had previously yielded 140 indexed lattices. Additionally, in tests onexperimental serial crystallography diffraction data recorded with quasi-monochromatic X-rays and with electrons the algorithm indexed more patternsthan other programs tested. more...
- Published
- 2020
14. Femtosecond phase-transition in hard x-ray excited bismuth
- Author
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Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., David, C., Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., and David, C. more...
- Abstract
The evolution of bismuth crystal structure upon excitation of its A(1g) phonon has been intensely studied with short pulse optical lasers. Here we present the first-time observation of a hard x-ray induced ultrafast phase transition in a bismuth single crystal at high intensities (similar to 10(14) W/cm(2)). The lattice evolution was followed using a recently demonstrated x-ray single-shot probing setup. The time evolution of the (111) Bragg peak intensity showed strong dependence on the excitation fluence. After exposure to a sufficiently intense x-ray pulse, the peak intensity dropped to zero within 300 fs, i.e. faster than one oscillation period of the A(1g) mode at room temperature. Our analysis indicates a nonthermal origin of a lattice disordering process, and excludes interpretations based on electron-ion equilibration process, or on thermodynamic heating process leading to plasma formation. more...
- Published
- 2019
- Full Text
- View/download PDF
15. Femtosecond phase-transition in hard x-ray excited bismuth
- Author
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Makita, M., Vartiainen, I., Mohacsi, I., Caleman, C., Diaz, A., Jönsson, Henrik Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., David, C., Makita, M., Vartiainen, I., Mohacsi, I., Caleman, C., Diaz, A., Jönsson, Henrik Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., and David, C. more...
- Abstract
The evolution of bismuth crystal structure upon excitation of its A(1g) phonon has been intensely studied with short pulse optical lasers. Here we present the first-time observation of a hard x-ray induced ultrafast phase transition in a bismuth single crystal at high intensities (similar to 10(14) W/cm(2)). The lattice evolution was followed using a recently demonstrated x-ray single-shot probing setup. The time evolution of the (111) Bragg peak intensity showed strong dependence on the excitation fluence. After exposure to a sufficiently intense x-ray pulse, the peak intensity dropped to zero within 300 fs, i.e. faster than one oscillation period of the A(1g) mode at room temperature. Our analysis indicates a nonthermal origin of a lattice disordering process, and excludes interpretations based on electron-ion equilibration process, or on thermodynamic heating process leading to plasma formation., QC 20190305 more...
- Published
- 2019
- Full Text
- View/download PDF
16. Femtosecond phase-transition in hard x-ray excited bismuth
- Author
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Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., David, C., Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., and David, C. more...
- Abstract
The evolution of bismuth crystal structure upon excitation of its A(1g) phonon has been intensely studied with short pulse optical lasers. Here we present the first-time observation of a hard x-ray induced ultrafast phase transition in a bismuth single crystal at high intensities (similar to 10(14) W/cm(2)). The lattice evolution was followed using a recently demonstrated x-ray single-shot probing setup. The time evolution of the (111) Bragg peak intensity showed strong dependence on the excitation fluence. After exposure to a sufficiently intense x-ray pulse, the peak intensity dropped to zero within 300 fs, i.e. faster than one oscillation period of the A(1g) mode at room temperature. Our analysis indicates a nonthermal origin of a lattice disordering process, and excludes interpretations based on electron-ion equilibration process, or on thermodynamic heating process leading to plasma formation. more...
- Published
- 2019
- Full Text
- View/download PDF
17. Femtosecond phase-transition in hard x-ray excited bismuth
- Author
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Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., David, C., Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., and David, C. more...
- Abstract
The evolution of bismuth crystal structure upon excitation of its A(1g) phonon has been intensely studied with short pulse optical lasers. Here we present the first-time observation of a hard x-ray induced ultrafast phase transition in a bismuth single crystal at high intensities (similar to 10(14) W/cm(2)). The lattice evolution was followed using a recently demonstrated x-ray single-shot probing setup. The time evolution of the (111) Bragg peak intensity showed strong dependence on the excitation fluence. After exposure to a sufficiently intense x-ray pulse, the peak intensity dropped to zero within 300 fs, i.e. faster than one oscillation period of the A(1g) mode at room temperature. Our analysis indicates a nonthermal origin of a lattice disordering process, and excludes interpretations based on electron-ion equilibration process, or on thermodynamic heating process leading to plasma formation. more...
- Published
- 2019
- Full Text
- View/download PDF
18. Femtosecond phase-transition in hard x-ray excited bismuth
- Author
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Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., David, C., Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., and David, C. more...
- Abstract
The evolution of bismuth crystal structure upon excitation of its A(1g) phonon has been intensely studied with short pulse optical lasers. Here we present the first-time observation of a hard x-ray induced ultrafast phase transition in a bismuth single crystal at high intensities (similar to 10(14) W/cm(2)). The lattice evolution was followed using a recently demonstrated x-ray single-shot probing setup. The time evolution of the (111) Bragg peak intensity showed strong dependence on the excitation fluence. After exposure to a sufficiently intense x-ray pulse, the peak intensity dropped to zero within 300 fs, i.e. faster than one oscillation period of the A(1g) mode at room temperature. Our analysis indicates a nonthermal origin of a lattice disordering process, and excludes interpretations based on electron-ion equilibration process, or on thermodynamic heating process leading to plasma formation. more...
- Published
- 2019
- Full Text
- View/download PDF
19. Femtosecond phase-transition in hard x-ray excited bismuth
- Author
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Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., David, C., Makita, M., Vartiainen, I., Mohacsi, I., Caleman, Carl, Diaz, A., Jönsson, Olof, Juranic, P., Medvedev, N., Meents, A., Mozzanica, A., Opara, N. L., Padeste, C., Panneels, V., Saxena, V., Sikorski, M., Song, S., Vera, L., Willmott, P. R., Beaud, P., Milne, C. J., Ziaja-Motyka, B., and David, C. more...
- Abstract
The evolution of bismuth crystal structure upon excitation of its A(1g) phonon has been intensely studied with short pulse optical lasers. Here we present the first-time observation of a hard x-ray induced ultrafast phase transition in a bismuth single crystal at high intensities (similar to 10(14) W/cm(2)). The lattice evolution was followed using a recently demonstrated x-ray single-shot probing setup. The time evolution of the (111) Bragg peak intensity showed strong dependence on the excitation fluence. After exposure to a sufficiently intense x-ray pulse, the peak intensity dropped to zero within 300 fs, i.e. faster than one oscillation period of the A(1g) mode at room temperature. Our analysis indicates a nonthermal origin of a lattice disordering process, and excludes interpretations based on electron-ion equilibration process, or on thermodynamic heating process leading to plasma formation. more...
- Published
- 2019
- Full Text
- View/download PDF
20. A DuMond-type crystal spectrometer for synchrotron-based X-ray emission studies in the energy range of 15–26 keV
- Author
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Jagodziński, Paweł, Szlachetko, Jakub, Dousse, Jean-Claude, Hoszowska, Joanna, Szlachetko, Monika, Vogelsang, U., Banaś, Dariusz, Pakendorf, T., Meents, A., Bokhoven, Jeroen A. van, Kubala-Kukuś, Aldona, Pajek, Marek, Nachtegaal, Maarten, Jagodziński, Paweł, Szlachetko, Jakub, Dousse, Jean-Claude, Hoszowska, Joanna, Szlachetko, Monika, Vogelsang, U., Banaś, Dariusz, Pakendorf, T., Meents, A., Bokhoven, Jeroen A. van, Kubala-Kukuś, Aldona, Pajek, Marek, and Nachtegaal, Maarten more...
- Abstract
The design and performance of a high-resolution transmission-type X-ray spectrometer for use in the 15–26 keV energy range at synchrotron light sources is reported. Monte Carlo X-ray-tracing simulations were performed to optimize the performance of the transmission-type spectrometer, based on the DuMond geometry, for use at the Super X-ray absorption beamline of the Swiss Light Source at the Paul Scherrer Institute. This spectrometer provides an instrumental energy resolution of 3.5 eV for X-ray emission lines around 16 keV and 12.5 eV for emission lines at 26 keV, which is comparable to the natural linewidths of the K and L X-ray transitions in the covered energy range. First experimental data are presented and compared with results of the Monte Carlo X-ray simulations. more...
- Published
- 2019
21. Transferring the entatic-state principle to copper photochemistry
- Author
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Dicke, B., Hoffmann, A., Stanek, J., Rampp, M. S., Grimm-Lebsanft, B., Biebl, F., Rukser, D., Maerz, B., Goeries, D., Naumova, M., Biednov, M., Neuber, G., Wetzel, A., Hofmann, S. M., Roedig, P., Meents, A., Bielecki, Johan, Andreasson, Jakob, Beyerlein, K. R., Chapman, H. N., Bressler, C., Zinth, W., Rübhausen, M., Herres-Pawlis, S., Dicke, B., Hoffmann, A., Stanek, J., Rampp, M. S., Grimm-Lebsanft, B., Biebl, F., Rukser, D., Maerz, B., Goeries, D., Naumova, M., Biednov, M., Neuber, G., Wetzel, A., Hofmann, S. M., Roedig, P., Meents, A., Bielecki, Johan, Andreasson, Jakob, Beyerlein, K. R., Chapman, H. N., Bressler, C., Zinth, W., Rübhausen, M., and Herres-Pawlis, S. more...
- Abstract
The entatic state denotes a distorted coordination geometry of a complex from its typical arrangement that generates an improvement to its function. The entatic-state principle has been observed to apply to copper electron-transfer proteins and it results in a lowering of the reorganization energy of the electron-transfer process. It is thus crucial for a multitude of biochemical processes, but its importance to photoactive complexes is unexplored. Here we study a copper complex-with a specifically designed constraining ligand geometry-that exhibits metal-to-ligand charge-transfer state lifetimes that are very short. The guanidine-quinoline ligand used here acts on the bis(chelated) copper(I) centre, allowing only small structural changes after photoexcitation that result in very fast structural dynamics. The data were collected using a multimethod approach that featured time-resolved ultraviolet-visible, infrared and X-ray absorption and optical emission spectroscopy. Through supporting density functional calculations, we deliver a detailed picture of the structural dynamics in the picosecond-to-nanosecond time range. more...
- Published
- 2018
- Full Text
- View/download PDF
22. X-ray focusing with efficient high-NA multilayer Laue lenses
- Author
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Bajt, S, Prasciolu, M, Fleckenstein, H, Domaracky, M, Chapman, HN, Morgan, AJ, Yefanov, O, Messerschmidt, M, Du, Y, Murray, KT, Mariani, V, Kuhn, M, Aplin, S, Pande, K, Villanueva-Perez, P, Stachnik, K, Chen, JPJ, Andrejczuk, A, Meents, A, Burkhardt, A, Pennicard, D, Huang, X, Yan, H, Nazaretski, E, Chu, YS, Hamm, CE, Bajt, S, Prasciolu, M, Fleckenstein, H, Domaracky, M, Chapman, HN, Morgan, AJ, Yefanov, O, Messerschmidt, M, Du, Y, Murray, KT, Mariani, V, Kuhn, M, Aplin, S, Pande, K, Villanueva-Perez, P, Stachnik, K, Chen, JPJ, Andrejczuk, A, Meents, A, Burkhardt, A, Pennicard, D, Huang, X, Yan, H, Nazaretski, E, Chu, YS, and Hamm, CE more...
- Abstract
Multilayer Laue lenses are volume diffraction elements for the efficient focusing of X-rays. With a new manufacturing technique that we introduced, it is possible to fabricate lenses of sufficiently high numerical aperture (NA) to achieve focal spot sizes below 10 nm. The alternating layers of the materials that form the lens must span a broad range of thicknesses on the nanometer scale to achieve the necessary range of X-ray deflection angles required to achieve a high NA. This poses a challenge to both the accuracy of the deposition process and the control of the materials properties, which often vary with layer thickness. We introduced a new pair of materials-tungsten carbide and silicon carbide-to prepare layered structures with smooth and sharp interfaces and with no material phase transitions that hampered the manufacture of previous lenses. Using a pair of multilayer Laue lenses (MLLs) fabricated from this system, we achieved a two-dimensional focus of 8.4 × 6.8 nm2 at a photon energy of 16.3 keV with high diffraction efficiency and demonstrated scanning-based imaging of samples with a resolution well below 10 nm. The high NA also allowed projection holographic imaging with strong phase contrast over a large range of magnifications. An error analysis indicates the possibility of achieving 1 nm focusing. more...
- Published
- 2018
23. Femtosecond X-ray coherent diffraction of aligned amyloid fibrils on low background graphene.
- Author
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Seuring, Carolin, Seuring, Carolin, Ayyer, Kartik, Filippaki, Eleftheria, Barthelmess, Miriam, Longchamp, Jean-Nicolas, Ringler, Philippe, Pardini, Tommaso, Wojtas, David H, Coleman, Matthew A, Dörner, Katerina, Fuglerud, Silje, Hammarin, Greger, Habenstein, Birgit, Langkilde, Annette E, Loquet, Antoine, Meents, Alke, Riek, Roland, Stahlberg, Henning, Boutet, Sébastien, Hunter, Mark S, Koglin, Jason, Liang, Mengning, Ginn, Helen M, Millane, Rick P, Frank, Matthias, Barty, Anton, Chapman, Henry N, Seuring, Carolin, Seuring, Carolin, Ayyer, Kartik, Filippaki, Eleftheria, Barthelmess, Miriam, Longchamp, Jean-Nicolas, Ringler, Philippe, Pardini, Tommaso, Wojtas, David H, Coleman, Matthew A, Dörner, Katerina, Fuglerud, Silje, Hammarin, Greger, Habenstein, Birgit, Langkilde, Annette E, Loquet, Antoine, Meents, Alke, Riek, Roland, Stahlberg, Henning, Boutet, Sébastien, Hunter, Mark S, Koglin, Jason, Liang, Mengning, Ginn, Helen M, Millane, Rick P, Frank, Matthias, Barty, Anton, and Chapman, Henry N more...
- Abstract
Here we present a new approach to diffraction imaging of amyloid fibrils, combining a free-standing graphene support and single nanofocused X-ray pulses of femtosecond duration from an X-ray free-electron laser. Due to the very low background scattering from the graphene support and mutual alignment of filaments, diffraction from tobacco mosaic virus (TMV) filaments and amyloid protofibrils is obtained to 2.7 Å and 2.4 Å resolution in single diffraction patterns, respectively. Some TMV diffraction patterns exhibit asymmetry that indicates the presence of a limited number of axial rotations in the XFEL focus. Signal-to-noise levels from individual diffraction patterns are enhanced using computational alignment and merging, giving patterns that are superior to those obtainable from synchrotron radiation sources. We anticipate that our approach will be a starting point for further investigations into unsolved structures of filaments and other weakly scattering objects. more...
- Published
- 2018
24. Femtosecond X-ray coherent diffraction of aligned amyloid fibrils on low background graphene
- Author
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Seuring, Carolin, Ayyer, Kartik, Filippaki, Eleftheria, Barthelmess, Miriam, Longchamp, Jean-Nicolas, Ringler, Philippe, Pardini, Tommaso, Wojtas, David H, Coleman, Matthew A, Dörner, Katerina, Fuglerud, Silje, Hammarin, Greger, Habenstein, Birgit, Langkilde, Annette E, Loquet, Antoine, Meents, Alke, Riek, Roland, Stahlberg, Henning, Boutet, Sébastien, Hunter, Mark S, Koglin, Jason, Liang, Mengning, Ginn, Helen M, Millane, Rick P, Frank, Matthias, Barty, Anton, Chapman, Henry N, Seuring, Carolin, Ayyer, Kartik, Filippaki, Eleftheria, Barthelmess, Miriam, Longchamp, Jean-Nicolas, Ringler, Philippe, Pardini, Tommaso, Wojtas, David H, Coleman, Matthew A, Dörner, Katerina, Fuglerud, Silje, Hammarin, Greger, Habenstein, Birgit, Langkilde, Annette E, Loquet, Antoine, Meents, Alke, Riek, Roland, Stahlberg, Henning, Boutet, Sébastien, Hunter, Mark S, Koglin, Jason, Liang, Mengning, Ginn, Helen M, Millane, Rick P, Frank, Matthias, Barty, Anton, and Chapman, Henry N more...
- Abstract
Here we present a new approach to diffraction imaging of amyloid fibrils, combining a free-standing graphene support and single nanofocused X-ray pulses of femtosecond duration from an X-ray free-electron laser. Due to the very low background scattering from the graphene support and mutual alignment of filaments, diffraction from tobacco mosaic virus (TMV) filaments and amyloid protofibrils is obtained to 2.7 Å and 2.4 Å resolution in single diffraction patterns, respectively. Some TMV diffraction patterns exhibit asymmetry that indicates the presence of a limited number of axial rotations in the XFEL focus. Signal-to-noise levels from individual diffraction patterns are enhanced using computational alignment and merging, giving patterns that are superior to those obtainable from synchrotron radiation sources. We anticipate that our approach will be a starting point for further investigations into unsolved structures of filaments and other weakly scattering objects. more...
- Published
- 2018
25. Femtosecond X-ray coherent diffraction of aligned amyloid fibrils on low background graphene
- Author
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Seuring, Carolin, Ayyer, Kartik, Filippaki, Eleftheria, Barthelmess, Miriam, Longchamp, Jean-Nicolas, Ringler, Philippe, Pardini, Tommaso, Wojtas, David H, Coleman, Matthew A, Dörner, Katerina, Fuglerud, Silje, Hammarin, Greger, Habenstein, Birgit, Langkilde, Annette E, Loquet, Antoine, Meents, Alke, Riek, Roland, Stahlberg, Henning, Boutet, Sébastien, Hunter, Mark S, Koglin, Jason, Liang, Mengning, Ginn, Helen M, Millane, Rick P, Frank, Matthias, Barty, Anton, Chapman, Henry N, Seuring, Carolin, Ayyer, Kartik, Filippaki, Eleftheria, Barthelmess, Miriam, Longchamp, Jean-Nicolas, Ringler, Philippe, Pardini, Tommaso, Wojtas, David H, Coleman, Matthew A, Dörner, Katerina, Fuglerud, Silje, Hammarin, Greger, Habenstein, Birgit, Langkilde, Annette E, Loquet, Antoine, Meents, Alke, Riek, Roland, Stahlberg, Henning, Boutet, Sébastien, Hunter, Mark S, Koglin, Jason, Liang, Mengning, Ginn, Helen M, Millane, Rick P, Frank, Matthias, Barty, Anton, and Chapman, Henry N more...
- Abstract
Here we present a new approach to diffraction imaging of amyloid fibrils, combining a free-standing graphene support and single nanofocused X-ray pulses of femtosecond duration from an X-ray free-electron laser. Due to the very low background scattering from the graphene support and mutual alignment of filaments, diffraction from tobacco mosaic virus (TMV) filaments and amyloid protofibrils is obtained to 2.7 Å and 2.4 Å resolution in single diffraction patterns, respectively. Some TMV diffraction patterns exhibit asymmetry that indicates the presence of a limited number of axial rotations in the XFEL focus. Signal-to-noise levels from individual diffraction patterns are enhanced using computational alignment and merging, giving patterns that are superior to those obtainable from synchrotron radiation sources. We anticipate that our approach will be a starting point for further investigations into unsolved structures of filaments and other weakly scattering objects. more...
- Published
- 2018
26. Mix-and-diffuse serial synchrotron crystallography
- Author
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Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, Oberthuer, Dominik, Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik more...
- Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources. more...
- Published
- 2017
- Full Text
- View/download PDF
27. Mix-and-diffuse serial synchrotron crystallography
- Author
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Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, Oberthuer, Dominik, Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik more...
- Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources. more...
- Published
- 2017
- Full Text
- View/download PDF
28. Mix-and-diffuse serial synchrotron crystallography
- Author
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Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, Oberthuer, Dominik, Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik more...
- Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources. more...
- Published
- 2017
- Full Text
- View/download PDF
29. High-speed fixed-target serial virus crystallography.
- Author
-
Roedig, Philip, Roedig, Philip, Ginn, Helen M, Pakendorf, Tim, Sutton, Geoff, Harlos, Karl, Walter, Thomas S, Meyer, Jan, Fischer, Pontus, Duman, Ramona, Vartiainen, Ismo, Reime, Bernd, Warmer, Martin, Brewster, Aaron S, Young, Iris D, Michels-Clark, Tara, Sauter, Nicholas K, Kotecha, Abhay, Kelly, James, Rowlands, David J, Sikorsky, Marcin, Nelson, Silke, Damiani, Daniel S, Alonso-Mori, Roberto, Ren, Jingshan, Fry, Elizabeth E, David, Christian, Stuart, David I, Wagner, Armin, Meents, Alke, Roedig, Philip, Roedig, Philip, Ginn, Helen M, Pakendorf, Tim, Sutton, Geoff, Harlos, Karl, Walter, Thomas S, Meyer, Jan, Fischer, Pontus, Duman, Ramona, Vartiainen, Ismo, Reime, Bernd, Warmer, Martin, Brewster, Aaron S, Young, Iris D, Michels-Clark, Tara, Sauter, Nicholas K, Kotecha, Abhay, Kelly, James, Rowlands, David J, Sikorsky, Marcin, Nelson, Silke, Damiani, Daniel S, Alonso-Mori, Roberto, Ren, Jingshan, Fry, Elizabeth E, David, Christian, Stuart, David I, Wagner, Armin, and Meents, Alke more...
- Abstract
We report a method for serial X-ray crystallography at X-ray free-electron lasers (XFELs), which allows for full use of the current 120-Hz repetition rate of the Linear Coherent Light Source (LCLS). Using a micropatterned silicon chip in combination with the high-speed Roadrunner goniometer for sample delivery, we were able to determine the crystal structures of the picornavirus bovine enterovirus 2 (BEV2) and the cytoplasmic polyhedrosis virus type 18 polyhedrin, with total data collection times of less than 14 and 10 min, respectively. Our method requires only micrograms of sample and should therefore broaden the applicability of serial femtosecond crystallography to challenging projects for which only limited sample amounts are available. By synchronizing the sample exchange to the XFEL repetition rate, our method allows for most efficient use of the limited beam time available at XFELs and should enable a substantial increase in sample throughput at these facilities. more...
- Published
- 2017
30. Mix-and-diffuse serial synchrotron crystallography
- Author
-
Beyerlein, KR, Dierksmeyer, D, Mariani, V, Kuhn, M, Sarrou, I, Ottaviano, A, Awel, S, Knoska, J, Fuglerud, S, Jonsson, O, Stern, S, Wiedorn, MO, Yefanov, O, Adriano, L, Bean, R, Burkhardt, A, Fischer, P, Heymann, M, Horke, DA, Jungnickel, KEJ, Kovaleva, E, Lorbeer, O, Metz, M, Meyer, J, Morgan, A, Pande, K, Panneerselvam, S, Seuring, C, Tolstikova, A, Lieske, J, Aplin, S, Roessle, M, White, TA, Chapman, HN, Meents, A, Oberthuer, D, Beyerlein, KR, Dierksmeyer, D, Mariani, V, Kuhn, M, Sarrou, I, Ottaviano, A, Awel, S, Knoska, J, Fuglerud, S, Jonsson, O, Stern, S, Wiedorn, MO, Yefanov, O, Adriano, L, Bean, R, Burkhardt, A, Fischer, P, Heymann, M, Horke, DA, Jungnickel, KEJ, Kovaleva, E, Lorbeer, O, Metz, M, Meyer, J, Morgan, A, Pande, K, Panneerselvam, S, Seuring, C, Tolstikova, A, Lieske, J, Aplin, S, Roessle, M, White, TA, Chapman, HN, Meents, A, and Oberthuer, D more...
- Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources. more...
- Published
- 2017
31. Structural enzymology using X-ray free electron lasers
- Author
-
Kupitz, C, Olmos, JL, Holl, M, Tremblay, L, Pande, K, Pandey, S, Oberthuer, D, Hunter, M, Liang, M, Aquila, A, Tenboer, J, Calvey, G, Katz, A, Chen, Y, Wiedorn, MO, Knoska, J, Meents, A, Majriani, V, Norwood, T, Poudyal, I, Grant, T, Miller, MD, Xu, W, Tolstikova, A, Morgan, A, Metz, M, Martin-Garcia, JM, Zook, JD, Roy-Chowdhury, S, Coe, J, Nagaratnam, N, Meza, D, Fromme, R, Basu, S, Frank, M, White, T, Barty, A, Bajt, S, Yefanov, O, Chapman, HN, Zatsepin, N, Nelson, G, Weierstall, U, Spence, J, Schwander, P, Pollack, L, Fromme, P, Ourmazd, A, Phillips, GN, Schmidt, M, Kupitz, C, Olmos, JL, Holl, M, Tremblay, L, Pande, K, Pandey, S, Oberthuer, D, Hunter, M, Liang, M, Aquila, A, Tenboer, J, Calvey, G, Katz, A, Chen, Y, Wiedorn, MO, Knoska, J, Meents, A, Majriani, V, Norwood, T, Poudyal, I, Grant, T, Miller, MD, Xu, W, Tolstikova, A, Morgan, A, Metz, M, Martin-Garcia, JM, Zook, JD, Roy-Chowdhury, S, Coe, J, Nagaratnam, N, Meza, D, Fromme, R, Basu, S, Frank, M, White, T, Barty, A, Bajt, S, Yefanov, O, Chapman, HN, Zatsepin, N, Nelson, G, Weierstall, U, Spence, J, Schwander, P, Pollack, L, Fromme, P, Ourmazd, A, Phillips, GN, and Schmidt, M more...
- Abstract
Mix-and-inject serial crystallography (MISC) is a technique designed to image enzyme catalyzed reactions in which small protein crystals are mixed with a substrate just prior to being probed by an X-ray pulse. This approach offers several advantages over flow cell studies. It provides (i) room temperature structures at near atomic resolution, (ii) time resolution ranging from microseconds to seconds, and (iii) convenient reaction initiation. It outruns radiation damage by using femtosecond X-ray pulses allowing damage and chemistry to be separated. Here, we demonstrate that MISC is feasible at an X-ray free electron laser by studying the reaction of M. tuberculosis ß-lactamase microcrystals with ceftriaxone antibiotic solution. Electron density maps of the apo-ß-lactamase and of the ceftriaxone bound form were obtained at 2.8 Å and 2.4 Å resolution, respectively. These results pave the way to study cyclic and non-cyclic reactions and represent a new field of time-resolved structural dynamics for numerous substrate-triggered biological reactions. more...
- Published
- 2017
32. High-speed fixed-target serial virus crystallography.
- Author
-
Roedig, Philip, Roedig, Philip, Ginn, Helen M, Pakendorf, Tim, Sutton, Geoff, Harlos, Karl, Walter, Thomas S, Meyer, Jan, Fischer, Pontus, Duman, Ramona, Vartiainen, Ismo, Reime, Bernd, Warmer, Martin, Brewster, Aaron S, Young, Iris D, Michels-Clark, Tara, Sauter, Nicholas K, Kotecha, Abhay, Kelly, James, Rowlands, David J, Sikorsky, Marcin, Nelson, Silke, Damiani, Daniel S, Alonso-Mori, Roberto, Ren, Jingshan, Fry, Elizabeth E, David, Christian, Stuart, David I, Wagner, Armin, Meents, Alke, Roedig, Philip, Roedig, Philip, Ginn, Helen M, Pakendorf, Tim, Sutton, Geoff, Harlos, Karl, Walter, Thomas S, Meyer, Jan, Fischer, Pontus, Duman, Ramona, Vartiainen, Ismo, Reime, Bernd, Warmer, Martin, Brewster, Aaron S, Young, Iris D, Michels-Clark, Tara, Sauter, Nicholas K, Kotecha, Abhay, Kelly, James, Rowlands, David J, Sikorsky, Marcin, Nelson, Silke, Damiani, Daniel S, Alonso-Mori, Roberto, Ren, Jingshan, Fry, Elizabeth E, David, Christian, Stuart, David I, Wagner, Armin, and Meents, Alke more...
- Abstract
We report a method for serial X-ray crystallography at X-ray free-electron lasers (XFELs), which allows for full use of the current 120-Hz repetition rate of the Linear Coherent Light Source (LCLS). Using a micropatterned silicon chip in combination with the high-speed Roadrunner goniometer for sample delivery, we were able to determine the crystal structures of the picornavirus bovine enterovirus 2 (BEV2) and the cytoplasmic polyhedrosis virus type 18 polyhedrin, with total data collection times of less than 14 and 10 min, respectively. Our method requires only micrograms of sample and should therefore broaden the applicability of serial femtosecond crystallography to challenging projects for which only limited sample amounts are available. By synchronizing the sample exchange to the XFEL repetition rate, our method allows for most efficient use of the limited beam time available at XFELs and should enable a substantial increase in sample throughput at these facilities. more...
- Published
- 2017
33. Structural enzymology using X-ray free electron lasers.
- Author
-
Kupitz, Christopher, Kupitz, Christopher, Olmos, Jose L, Holl, Mark, Tremblay, Lee, Pande, Kanupriya, Pandey, Suraj, Oberthür, Dominik, Hunter, Mark, Liang, Mengning, Aquila, Andrew, Tenboer, Jason, Calvey, George, Katz, Andrea, Chen, Yujie, Wiedorn, Max O, Knoska, Juraj, Meents, Alke, Majriani, Valerio, Norwood, Tyler, Poudyal, Ishwor, Grant, Thomas, Miller, Mitchell D, Xu, Weijun, Tolstikova, Aleksandra, Morgan, Andrew, Metz, Markus, Martin-Garcia, Jose M, Zook, James D, Roy-Chowdhury, Shatabdi, Coe, Jesse, Nagaratnam, Nirupa, Meza, Domingo, Fromme, Raimund, Basu, Shibom, Frank, Matthias, White, Thomas, Barty, Anton, Bajt, Sasa, Yefanov, Oleksandr, Chapman, Henry N, Zatsepin, Nadia, Nelson, Garrett, Weierstall, Uwe, Spence, John, Schwander, Peter, Pollack, Lois, Fromme, Petra, Ourmazd, Abbas, Phillips, George N, Schmidt, Marius, Kupitz, Christopher, Kupitz, Christopher, Olmos, Jose L, Holl, Mark, Tremblay, Lee, Pande, Kanupriya, Pandey, Suraj, Oberthür, Dominik, Hunter, Mark, Liang, Mengning, Aquila, Andrew, Tenboer, Jason, Calvey, George, Katz, Andrea, Chen, Yujie, Wiedorn, Max O, Knoska, Juraj, Meents, Alke, Majriani, Valerio, Norwood, Tyler, Poudyal, Ishwor, Grant, Thomas, Miller, Mitchell D, Xu, Weijun, Tolstikova, Aleksandra, Morgan, Andrew, Metz, Markus, Martin-Garcia, Jose M, Zook, James D, Roy-Chowdhury, Shatabdi, Coe, Jesse, Nagaratnam, Nirupa, Meza, Domingo, Fromme, Raimund, Basu, Shibom, Frank, Matthias, White, Thomas, Barty, Anton, Bajt, Sasa, Yefanov, Oleksandr, Chapman, Henry N, Zatsepin, Nadia, Nelson, Garrett, Weierstall, Uwe, Spence, John, Schwander, Peter, Pollack, Lois, Fromme, Petra, Ourmazd, Abbas, Phillips, George N, and Schmidt, Marius more...
- Abstract
Mix-and-inject serial crystallography (MISC) is a technique designed to image enzyme catalyzed reactions in which small protein crystals are mixed with a substrate just prior to being probed by an X-ray pulse. This approach offers several advantages over flow cell studies. It provides (i) room temperature structures at near atomic resolution, (ii) time resolution ranging from microseconds to seconds, and (iii) convenient reaction initiation. It outruns radiation damage by using femtosecond X-ray pulses allowing damage and chemistry to be separated. Here, we demonstrate that MISC is feasible at an X-ray free electron laser by studying the reaction of M. tuberculosis ß-lactamase microcrystals with ceftriaxone antibiotic solution. Electron density maps of the apo-ß-lactamase and of the ceftriaxone bound form were obtained at 2.8 Å and 2.4 Å resolution, respectively. These results pave the way to study cyclic and non-cyclic reactions and represent a new field of time-resolved structural dynamics for numerous substrate-triggered biological reactions. more...
- Published
- 2017
34. Mix-and-diffuse serial synchrotron crystallography
- Author
-
Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, Oberthuer, Dominik, Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik more...
- Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources. more...
- Published
- 2017
- Full Text
- View/download PDF
35. Mix-and-diffuse serial synchrotron crystallography
- Author
-
Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, Oberthuer, Dominik, Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik more...
- Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources. more...
- Published
- 2017
- Full Text
- View/download PDF
36. Early Point-of-Care Platelet Function Testing Using Multiple Electrode Aggregometry in Patients Undergoing Cardiac Surgery
- Author
-
Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, Noordzij, Peter G., Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, and Noordzij, Peter G. more...
- Published
- 2016
37. SCN10A Mutation in a Patient with Erythromelalgia Enhances C-Fiber Activity Dependent Slowing
- Author
-
Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, Namer, Barbara, Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, and Namer, Barbara more...
- Abstract
Gain-of-function mutations in the tetrodotoxin (TTX) sensitive voltage-gated sodium channel (Nav) Nav1.7 have been identified as a key mechanism underlying chronic pain in inherited erythromelalgia. Mutations in TTX resistant channels, such as Nav1.8 or Nav1.9, were recently connected with inherited chronic pain syndromes. Here, we investigated the effects of the p.M650K mutation in Nav1.8 in a 53 year old patient with erythromelalgia by micro-neurography and patch-clamp techniques. Recordings of the patient's peripheral nerve fibers showed increased activity dependent slowing (ADS) in CMi and less spontaneous firing compared to a control group of erythromelalgia patients without Nav mutations. To evaluate the impact of the p. M650K mutation on neuronal firing and channel gating, we performed current and voltage-clamp recordings on transfected sensory neurons (DRGs) and neuroblastoma cells. The p. M650K mutation shifted steady-state fast inactivation of Nav1.8 to more hyperpolarized potentials and did not significantly alter any other tested gating behaviors. The AP half-width was significantly broader and the stimulated action potential firing rate was reduced for M650K transfected DRGs compared to WT. We discuss the potential link between enhanced steady state fast inactivation, broader action potential width and the potential physiological consequences. more...
- Published
- 2016
- Full Text
- View/download PDF
38. SCN10A Mutation in a Patient with Erythromelalgia Enhances C-Fiber Activity Dependent Slowing
- Author
-
Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, Namer, Barbara, Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, and Namer, Barbara more...
- Abstract
Gain-of-function mutations in the tetrodotoxin (TTX) sensitive voltage-gated sodium channel (Nav) Nav1.7 have been identified as a key mechanism underlying chronic pain in inherited erythromelalgia. Mutations in TTX resistant channels, such as Nav1.8 or Nav1.9, were recently connected with inherited chronic pain syndromes. Here, we investigated the effects of the p.M650K mutation in Nav1.8 in a 53 year old patient with erythromelalgia by micro-neurography and patch-clamp techniques. Recordings of the patient's peripheral nerve fibers showed increased activity dependent slowing (ADS) in CMi and less spontaneous firing compared to a control group of erythromelalgia patients without Nav mutations. To evaluate the impact of the p. M650K mutation on neuronal firing and channel gating, we performed current and voltage-clamp recordings on transfected sensory neurons (DRGs) and neuroblastoma cells. The p. M650K mutation shifted steady-state fast inactivation of Nav1.8 to more hyperpolarized potentials and did not significantly alter any other tested gating behaviors. The AP half-width was significantly broader and the stimulated action potential firing rate was reduced for M650K transfected DRGs compared to WT. We discuss the potential link between enhanced steady state fast inactivation, broader action potential width and the potential physiological consequences. more...
- Published
- 2016
- Full Text
- View/download PDF
39. SCN10A Mutation in a Patient with Erythromelalgia Enhances C-Fiber Activity Dependent Slowing
- Author
-
Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, Namer, Barbara, Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, and Namer, Barbara more...
- Abstract
Gain-of-function mutations in the tetrodotoxin (TTX) sensitive voltage-gated sodium channel (Nav) Nav1.7 have been identified as a key mechanism underlying chronic pain in inherited erythromelalgia. Mutations in TTX resistant channels, such as Nav1.8 or Nav1.9, were recently connected with inherited chronic pain syndromes. Here, we investigated the effects of the p.M650K mutation in Nav1.8 in a 53 year old patient with erythromelalgia by micro-neurography and patch-clamp techniques. Recordings of the patient's peripheral nerve fibers showed increased activity dependent slowing (ADS) in CMi and less spontaneous firing compared to a control group of erythromelalgia patients without Nav mutations. To evaluate the impact of the p. M650K mutation on neuronal firing and channel gating, we performed current and voltage-clamp recordings on transfected sensory neurons (DRGs) and neuroblastoma cells. The p. M650K mutation shifted steady-state fast inactivation of Nav1.8 to more hyperpolarized potentials and did not significantly alter any other tested gating behaviors. The AP half-width was significantly broader and the stimulated action potential firing rate was reduced for M650K transfected DRGs compared to WT. We discuss the potential link between enhanced steady state fast inactivation, broader action potential width and the potential physiological consequences. more...
- Published
- 2016
- Full Text
- View/download PDF
40. TakeTwo: an indexing algorithm suited to still images with known crystal parameters.
- Author
-
Ginn, Helen Mary, Ginn, Helen Mary, Roedig, Philip, Kuo, Anling, Evans, Gwyndaf, Sauter, Nicholas K, Ernst, Oliver P, Meents, Alke, Mueller-Werkmeister, Henrike, Miller, RJ Dwayne, Stuart, David Ian, Ginn, Helen Mary, Ginn, Helen Mary, Roedig, Philip, Kuo, Anling, Evans, Gwyndaf, Sauter, Nicholas K, Ernst, Oliver P, Meents, Alke, Mueller-Werkmeister, Henrike, Miller, RJ Dwayne, and Stuart, David Ian more...
- Abstract
The indexing methods currently used for serial femtosecond crystallography were originally developed for experiments in which crystals are rotated in the X-ray beam, providing significant three-dimensional information. On the other hand, shots from both X-ray free-electron lasers and serial synchrotron crystallography experiments are still images, in which the few three-dimensional data available arise only from the curvature of the Ewald sphere. Traditional synchrotron crystallography methods are thus less well suited to still image data processing. Here, a new indexing method is presented with the aim of maximizing information use from a still image given the known unit-cell dimensions and space group. Efficacy for cubic, hexagonal and orthorhombic space groups is shown, and for those showing some evidence of diffraction the indexing rate ranged from 90% (hexagonal space group) to 151% (cubic space group). Here, the indexing rate refers to the number of lattices indexed per image. more...
- Published
- 2016
41. § 15 Die internationale Zuständigkeit bei grenzüberschreitenden Rechtsstreitigkeiten über Cloud Computing
- Author
-
Borges, Georg, Meents, Jan Geert, Thole, Christoph, Borges, Georg, Meents, Jan Geert, and Thole, Christoph
- Published
- 2016
42. Early Point-of-Care Platelet Function Testing Using Multiple Electrode Aggregometry in Patients Undergoing Cardiac Surgery
- Author
-
Sub Pharmacotherapy, Theoretical, Pharmacoepidemiology and Clinical Pharmacology, Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, Noordzij, Peter G., Sub Pharmacotherapy, Theoretical, Pharmacoepidemiology and Clinical Pharmacology, Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, and Noordzij, Peter G. more...
- Published
- 2016
43. Free-Flow Electrophoresis of Plasma Membrane Vesicles Enriched by Two-Phase Partitioning Enhances the Quality of the Proteome from Arabidopsis Seedlings
- Author
-
de Michele, R, McFarlane, HE, Parsons, HT, Meents, MJ, Lao, J, Fernandez-Nino, SMG, Petzold, CJ, Frommer, WB, Samuels, AL, Heazlewood, JL, de Michele, R, McFarlane, HE, Parsons, HT, Meents, MJ, Lao, J, Fernandez-Nino, SMG, Petzold, CJ, Frommer, WB, Samuels, AL, and Heazlewood, JL more...
- Abstract
The plant plasma membrane is the interface between the cell and its environment undertaking a range of important functions related to transport, signaling, cell wall biosynthesis, and secretion. Multiple proteomic studies have attempted to capture the diversity of proteins in the plasma membrane using biochemical fractionation techniques. In this study, two-phase partitioning was combined with free-flow electrophoresis to produce a population of highly purified plasma membrane vesicles that were subsequently characterized by tandem mass spectroscopy. This combined high-quality plasma membrane isolation technique produced a reproducible proteomic library of over 1000 proteins with an extended dynamic range including plasma membrane-associated proteins. The approach enabled the detection of a number of putative plasma membrane proteins not previously identified by other studies, including peripheral membrane proteins. Utilizing multiple data sources, we developed a PM-confidence score to provide a value indicating association to the plasma membrane. This study highlights over 700 proteins that, while seemingly abundant at the plasma membrane, are mostly unstudied. To validate this data set, we selected 14 candidates and transiently localized 13 to the plasma membrane using a fluorescent tag. Given the importance of the plasma membrane, this data set provides a valuable tool to further investigate important proteins. The mass spectrometry data are available via ProteomeXchange, identifier PXD001795. more...
- Published
- 2016
44. Early Point-of-Care Platelet Function Testing Using Multiple Electrode Aggregometry in Patients Undergoing Cardiac Surgery
- Author
-
Sub Pharmacotherapy, Theoretical, Pharmacoepidemiology and Clinical Pharmacology, Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, Noordzij, Peter G., Sub Pharmacotherapy, Theoretical, Pharmacoepidemiology and Clinical Pharmacology, Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, and Noordzij, Peter G. more...
- Published
- 2016
45. TakeTwo: an indexing algorithm suited to still images with known crystal parameters.
- Author
-
Ginn, Helen Mary, Ginn, Helen Mary, Roedig, Philip, Kuo, Anling, Evans, Gwyndaf, Sauter, Nicholas K, Ernst, Oliver P, Meents, Alke, Mueller-Werkmeister, Henrike, Miller, RJ Dwayne, Stuart, David Ian, Ginn, Helen Mary, Ginn, Helen Mary, Roedig, Philip, Kuo, Anling, Evans, Gwyndaf, Sauter, Nicholas K, Ernst, Oliver P, Meents, Alke, Mueller-Werkmeister, Henrike, Miller, RJ Dwayne, and Stuart, David Ian more...
- Abstract
The indexing methods currently used for serial femtosecond crystallography were originally developed for experiments in which crystals are rotated in the X-ray beam, providing significant three-dimensional information. On the other hand, shots from both X-ray free-electron lasers and serial synchrotron crystallography experiments are still images, in which the few three-dimensional data available arise only from the curvature of the Ewald sphere. Traditional synchrotron crystallography methods are thus less well suited to still image data processing. Here, a new indexing method is presented with the aim of maximizing information use from a still image given the known unit-cell dimensions and space group. Efficacy for cubic, hexagonal and orthorhombic space groups is shown, and for those showing some evidence of diffraction the indexing rate ranged from 90% (hexagonal space group) to 151% (cubic space group). Here, the indexing rate refers to the number of lattices indexed per image. more...
- Published
- 2016
46. AXSIS: Exploring the frontiers in attosecond X-ray science, imaging and spectroscopy.
- Author
-
Kärtner, FX, Kärtner, FX, Ahr, F, Calendron, A-L, Çankaya, H, Carbajo, S, Chang, G, Cirmi, G, Dörner, K, Dorda, U, Fallahi, A, Hartin, A, Hemmer, M, Hobbs, R, Hua, Y, Huang, WR, Letrun, R, Matlis, N, Mazalova, V, Mücke, OD, Nanni, E, Putnam, W, Ravi, K, Reichert, F, Sarrou, I, Wu, X, Yahaghi, A, Ye, H, Zapata, L, Zhang, D, Zhou, C, Miller, RJD, Berggren, KK, Graafsma, H, Meents, A, Assmann, RW, Chapman, HN, Fromme, P, Kärtner, FX, Kärtner, FX, Ahr, F, Calendron, A-L, Çankaya, H, Carbajo, S, Chang, G, Cirmi, G, Dörner, K, Dorda, U, Fallahi, A, Hartin, A, Hemmer, M, Hobbs, R, Hua, Y, Huang, WR, Letrun, R, Matlis, N, Mazalova, V, Mücke, OD, Nanni, E, Putnam, W, Ravi, K, Reichert, F, Sarrou, I, Wu, X, Yahaghi, A, Ye, H, Zapata, L, Zhang, D, Zhou, C, Miller, RJD, Berggren, KK, Graafsma, H, Meents, A, Assmann, RW, Chapman, HN, and Fromme, P more...
- Abstract
X-ray crystallography is one of the main methods to determine atomic-resolution 3D images of the whole spectrum of molecules ranging from small inorganic clusters to large protein complexes consisting of hundred-thousands of atoms that constitute the macromolecular machinery of life. Life is not static, and unravelling the structure and dynamics of the most important reactions in chemistry and biology is essential to uncover their mechanism. Many of these reactions, including photosynthesis which drives our biosphere, are light induced and occur on ultrafast timescales. These have been studied with high time resolution primarily by optical spectroscopy, enabled by ultrafast laser technology, but they reduce the vast complexity of the process to a few reaction coordinates. In the AXSIS project at CFEL in Hamburg, funded by the European Research Council, we develop the new method of attosecond serial X-ray crystallography and spectroscopy, to give a full description of ultrafast processes atomically resolved in real space and on the electronic energy landscape, from co-measurement of X-ray and optical spectra, and X-ray diffraction. This technique will revolutionize our understanding of structure and function at the atomic and molecular level and thereby unravel fundamental processes in chemistry and biology like energy conversion processes. For that purpose, we develop a compact, fully coherent, THz-driven atto-second X-ray source based on coherent inverse Compton scattering off a free-electron crystal, to outrun radiation damage effects due to the necessary high X-ray irradiance required to acquire diffraction signals. This highly synergistic project starts from a completely clean slate rather than conforming to the specifications of a large free-electron laser (FEL) user facility, to optimize the entire instrumentation towards fundamental measurements of the mechanism of light absorption and excitation energy transfer. A multidisciplinary team formed by laser-, acc more...
- Published
- 2016
47. SCN10A Mutation in a Patient with Erythromelalgia Enhances C-Fiber Activity Dependent Slowing
- Author
-
Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, Namer, Barbara, Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, and Namer, Barbara more...
- Abstract
Gain-of-function mutations in the tetrodotoxin (TTX) sensitive voltage-gated sodium channel (Nav) Nav1.7 have been identified as a key mechanism underlying chronic pain in inherited erythromelalgia. Mutations in TTX resistant channels, such as Nav1.8 or Nav1.9, were recently connected with inherited chronic pain syndromes. Here, we investigated the effects of the p.M650K mutation in Nav1.8 in a 53 year old patient with erythromelalgia by micro-neurography and patch-clamp techniques. Recordings of the patient's peripheral nerve fibers showed increased activity dependent slowing (ADS) in CMi and less spontaneous firing compared to a control group of erythromelalgia patients without Nav mutations. To evaluate the impact of the p. M650K mutation on neuronal firing and channel gating, we performed current and voltage-clamp recordings on transfected sensory neurons (DRGs) and neuroblastoma cells. The p. M650K mutation shifted steady-state fast inactivation of Nav1.8 to more hyperpolarized potentials and did not significantly alter any other tested gating behaviors. The AP half-width was significantly broader and the stimulated action potential firing rate was reduced for M650K transfected DRGs compared to WT. We discuss the potential link between enhanced steady state fast inactivation, broader action potential width and the potential physiological consequences. more...
- Published
- 2016
- Full Text
- View/download PDF
48. Early Point-of-Care Platelet Function Testing Using Multiple Electrode Aggregometry in Patients Undergoing Cardiac Surgery
- Author
-
Sub Pharmacotherapy, Theoretical, Pharmacoepidemiology and Clinical Pharmacology, Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, Noordzij, Peter G., Sub Pharmacotherapy, Theoretical, Pharmacoepidemiology and Clinical Pharmacology, Vlot, Eline A., Meents, Nanda, van de Garde, Ewoudt M.W., Hackeng, Christian M, and Noordzij, Peter G. more...
- Published
- 2016
49. § 15 Die internationale Zuständigkeit bei grenzüberschreitenden Rechtsstreitigkeiten über Cloud Computing
- Author
-
Borges, Georg, Meents, Jan Geert, Thole, Christoph, Borges, Georg, Meents, Jan Geert, and Thole, Christoph
- Published
- 2016
50. SCN10A Mutation in a Patient with Erythromelalgia Enhances C-Fiber Activity Dependent Slowing
- Author
-
Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, Namer, Barbara, Kist, Andreas M., Sagafos, Dagrun, Rush, Anthony M., Neacsu, Cristian, Eberhardt, Esther, Schmidt, Roland, Lunden, Lars Kristian, Orstavik, Kristin, Kaluza, Luisa, Meents, Jannis, Zhang, Zhiping, Carr, Thomas Hedley, Salter, Hugh, Malinowsky, David, Wollberg, Patrik, Krupp, Johannes, Kleggetveit, Inge Petter, Schmelz, Martin, Jorum, Ellen, Lampert, Angelika, and Namer, Barbara more...
- Abstract
Gain-of-function mutations in the tetrodotoxin (TTX) sensitive voltage-gated sodium channel (Nav) Nav1.7 have been identified as a key mechanism underlying chronic pain in inherited erythromelalgia. Mutations in TTX resistant channels, such as Nav1.8 or Nav1.9, were recently connected with inherited chronic pain syndromes. Here, we investigated the effects of the p.M650K mutation in Nav1.8 in a 53 year old patient with erythromelalgia by micro-neurography and patch-clamp techniques. Recordings of the patient's peripheral nerve fibers showed increased activity dependent slowing (ADS) in CMi and less spontaneous firing compared to a control group of erythromelalgia patients without Nav mutations. To evaluate the impact of the p. M650K mutation on neuronal firing and channel gating, we performed current and voltage-clamp recordings on transfected sensory neurons (DRGs) and neuroblastoma cells. The p. M650K mutation shifted steady-state fast inactivation of Nav1.8 to more hyperpolarized potentials and did not significantly alter any other tested gating behaviors. The AP half-width was significantly broader and the stimulated action potential firing rate was reduced for M650K transfected DRGs compared to WT. We discuss the potential link between enhanced steady state fast inactivation, broader action potential width and the potential physiological consequences. more...
- Published
- 2016
- Full Text
- View/download PDF
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