4 results on '"Agata Rekas"'
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2. Robust High-Yield Methodologies for 2H and 2H/15N/13C Labeling of Proteins for Structural Investigations Using Neutron Scattering and NMR
- Author
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Peter J. Holden, Agata Rekas, Anthony P. Duff, Vanessa Lake, and Karyn L Wilde
- Subjects
chemistry.chemical_compound ,Protein structure ,Deuterium ,Exponential growth ,Chemistry ,Analytical chemistry ,medicine ,Glycerol ,Neutron ,Nuclear magnetic resonance spectroscopy ,Neutron scattering ,medicine.disease_cause ,Escherichia coli - Abstract
We have developed a method that has proven highly reliable for the deuteration and triple labeling ((2)H/(15)N/(13)C) of a broad range of proteins by recombinant expression in Escherichia coli BL21. Typical biomass yields are 40-80g/L wet weight, yielding 50-500mg/L purified protein. This method uses a simple, relatively inexpensive defined medium, and routinely results in a high-yield expression without need for optimization. The key elements are very tight control of expression, careful starter culture adaptation steps, media composition, and strict maintenance of aerobic conditions ensuring exponential growth. Temperature is reduced as required to prevent biological oxygen demand exceeding maximum aeration capacity. Glycerol is the sole carbon source. We have not encountered an upper limit for the size of proteins that can be expressed, achieving excellent expression for proteins from 11 to 154kDa and the quantity produced at 1L scale ensures that no small-angle neutron scattering, nuclear magnetic resonance, or neutron crystallography experiment is limited by the amount of deuterated material. Where difficulties remain, these tend to be cases of altered protein solubility due to high protein concentration and a D2O-based environment.
- Published
- 2015
- Full Text
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3. Dopamine-Induced α-Synuclein Oligomers
- Author
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Agata Rekas, Roberto Cappai, Chi L.L. Pham, and Cyril C. Curtain
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Parkinson's disease ,Amyloid ,Pars compacta ,Dementia with Lewy bodies ,Dopaminergic ,Substantia nigra ,medicine.disease ,nervous system diseases ,Cell biology ,chemistry.chemical_compound ,nervous system ,chemistry ,Dopamine ,medicine ,Neurotransmitter ,Neuroscience ,medicine.drug - Abstract
α-Synuclein (α-syn) is a 140-residue natively unfolded protein; its physiologic role is not yet elucidated, although it has been implicated in a number of neurodegenerative disorders, including Parkinson’s disease (PD), dementia with Lewy bodies (LB) and the LB variant of Parkinson’s disease. PD is characterized by a selective loss of dopaminergic neurons and the presence of insoluble α-syn aggregates, i.e. amyloid fibrils, deposited in LBs in the remaining substantia nigra pars compacta neurons of the mid-brain. This region contains neurons whose main neurotransmitter is dopamine (DA) and thus plays a role in reward-driven learning and control of movement and balance. This chapter focuses on the interaction of α-syn and DA, in particular on the resulting soluble oligomers, their structure, properties and potential role in the development and prevention of PD.
- Published
- 2014
- Full Text
- View/download PDF
4. Contributors
- Author
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Marie-Isabel Aguilar, Maria Andreasen, Neil R. Anthony, Fernando Teran Arce, Florian Arnhold, Shruti Arya, Aida Attar, Silvia Barbosa, Manja A. Behrens, Innocent B. Bekard, Georges Belfort, Martin L. Bennink, Magdalena Bereza, Keith M. Berland, Gal Bitan, Mischa Bonn, Hugo M. Botelho, Joana Branco dos Santos, Leonid Breydo, Yi Cao, Roberto Cappai, Isabel Cardoso, Mariano Carrión-Vázquez, Sofia B. Carvalho, John A. Carver, Fiona T.S. Chan, W. Seth Childers, Jason C. Collins, Laura Connelly, Cyril Curtain, Vijit Dalal, Alfonso De Simone, Athene M. Donald, Dave E. Dunstan, Heath Ecroyd, Jan Johannes Enghild, Laura Esther Abelleira, Marcus Fändrich, María del Carmen Fernández-Ramírez, Vito Foderà, Günter Fritz, Yoshiaki Furukawa, Shirin D. Ghodke, Stefano Gianni, Armin Giese, Cláudio M. Gomes, Susana Gonçalves, Lesley H. Greene, Michael D.W. Griffin, Amy M. Griggs, Christian Haass, Per Hammarström, J. Robin Harris, Federico Herrera, Rubén Hervás, Shannon E. Hill, Xu Hou, Geoffrey J. Howlett, Tsuneya Ikezu, Hyunbum Jang, Grethe V. Jensen, Bruce L. Kagan, Clemens F. Kaminski, Shigeko Kawai-Noma, Alexei Klechikov, Gijsje H. Koenderink, Ratnesh Lal, Douglas V. Laurents, Chi Le Lan Pham, Harry LeVine, Dusan Losic, David G. Lynn, Kirsten G. Malmos, Lisandra L. Martin, Adam I. Mechler, Anil K. Mehta, Derya Meral, Nathaniel G.N. Milton, Yee-Foong Mok, Ludmilla A. Morozova-Roche, Víctor Mosquera, Samrat Mukhopadhyay, Mentor Mulaj, Martin Muschol, Niels Chr Nielsen, R. Nilsson, Georg Nübling, Ruth Nussinov, Daniel E. Otzen, Tiago Fleming Outeiro, Jan Skov Pedersen, Jesper Søndergaard Pedersen, K. Peter, Dorothea Pinotsi, Meng Qin, Srinivasan Ramachandran, Agata Rekas, Annalisa Relini, Jean-Christophe Rochet, Sophie Rothhämel, Nikita Rudinskiy, Markus Sauer, Gabriele S. Kaminski Schierle, Michael Schleeger, Bettina Schmid, David H. Small, Anne Søndergaard, Mirco Sorci, Tara L. Spires-Jones, Marcel Stenvang, Martin Stöckl, Katrin Strecker, Vinod Subramaniam, Anna S.P. Svane, Kim K.M. Sweers, Pablo Taboada, Hideki Taguchi, Chai Lean Teoh, David C. Thorn, Kiyotaka Tokuraku, Robert Tycko, Brigita Urbanc, Vladimir N. Uversky, Corianne C. van den Akker, Michele Vendruscolo, Anna von Mikecz, Vladana Vukojević, Wei Wang, Katrin Weise, Roland Winter, Jessica W. Wu, Wei-Feng Xue, Kiran Yanamandra, Daniel Ysselstein, Eva Žerovnik, and Dawei Zou
- Published
- 2014
- Full Text
- View/download PDF
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