5 results on '"T, Kaufhold"'
Search Results
2. Repeat DNA-PAINT suppresses background and non-specific signals in optical nanoscopy
- Author
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Alexander H. Clowsley, William T. Kaufhold, Tobias Lutz, Anna Meletiou, Lorenzo Di Michele, and Christian Soeller
- Subjects
Science - Abstract
DNA-PAINT is a super-resolution imaging technique which suffers from high background signals and non-specific binding. Here the authors report Repeat DNA-PAINT which is capable of supressing background noise and preventing photoinduced site loss, as well as decreasing the time taken for the sampling process.
- Published
- 2021
- Full Text
- View/download PDF
3. Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni.
- Author
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Jagamya Vijayaraghavan, Vijay Kumar, Nikhil P Krishnan, Ross T Kaufhold, Ximin Zeng, Jun Lin, and Focco van den Akker
- Subjects
Medicine ,Science - Abstract
The bacterial soluble lytic transglycosylase (LT) breaks down the peptidoglycan (PG) layer during processes such as cell division. We present here crystal structures of the soluble LT Cj0843 from Campylobacter jejuni with and without bulgecin A inhibitor in the active site. Cj0843 has a doughnut shape similar but not identical to that of E. coli SLT70. The C-terminal catalytic domain is preceded by an L-domain, a large helical U-domain, a flexible linker, and a small N-terminal NU-domain. The flexible linker allows the NU-domain to reach over and complete the circular shape, using residues conserved in the Epsilonproteobacteria LT family. The inner surface of the Cj0843 doughnut is mostly positively charged including a pocket that has 8 Arg/Lys residues. Molecular dynamics simulations with PG strands revealed a potential functional role for this pocket in anchoring the negatively charged terminal tetrapeptide of the PG during several steps in the reaction including homing and aligning the PG strand for exolytic cleavage, and subsequent ratcheting of the PG strand to enhance processivity in degrading PG strands.
- Published
- 2018
- Full Text
- View/download PDF
4. Responsive core-shell DNA particles trigger lipid-membrane disruption and bacteria entrapment
- Author
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Lorenzo Di Michele, Ryan A. Brady, Roger Rubio-Sánchez, Pietro Cicuta, Michal Walczak, Leonardo Mancini, Claudia Contini, William T. Kaufhold, Walczak, Michal [0000-0002-4701-9476], Contini, Claudia [0000-0002-5282-2458], Rubio-Sánchez, Roger [0000-0001-5574-5809], Cicuta, Pietro [0000-0002-9193-8496], Di Michele, Lorenzo [0000-0002-1458-9747], Apollo - University of Cambridge Repository, Commission of the European Communities, and The Royal Society
- Subjects
Pore Forming Cytotoxic Proteins ,Cell Membrane Permeability ,Membrane permeability ,123 ,Science ,Antimicrobial peptides ,147 ,General Physics and Astronomy ,DNA nanostructures ,02 engineering and technology ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,chemistry.chemical_compound ,Membrane Lipids ,639/638/298/54/989 ,Escherichia coli ,Secretion ,128 ,Particle Size ,Lipid bilayer ,030304 developmental biology ,0303 health sciences ,Multidisciplinary ,132 ,biology ,Bacteria ,Organizing materials with DNA ,Vesicle ,Bioinspired materials ,article ,General Chemistry ,DNA ,021001 nanoscience & nanotechnology ,biology.organism_classification ,639/925/926/1050 ,3. Good health ,Nanostructures ,Membrane ,chemistry ,Biophysics ,639/925/926/1049 ,0210 nano-technology ,147/143 ,Hydrophobic and Hydrophilic Interactions - Abstract
Biology has evolved a variety of agents capable of permeabilizing and disrupting lipid membranes, from amyloid aggregates, to antimicrobial peptides, to venom compounds. While often associated with disease or toxicity, these agents are also central to many biosensing and therapeutic technologies. Here, we introduce a class of synthetic, DNA-based particles capable of disrupting lipid membranes. The particles have finely programmable size, and self-assemble from all-DNA and cholesterol-DNA nanostructures, the latter forming a membrane-adhesive core and the former a protective hydrophilic corona. We show that the corona can be selectively displaced with a molecular cue, exposing the ‘sticky’ core. Unprotected particles adhere to synthetic lipid vesicles, which in turn enhances membrane permeability and leads to vesicle collapse. Furthermore, particle-particle coalescence leads to the formation of gel-like DNA aggregates that envelop surviving vesicles. This response is reminiscent of pathogen immobilisation through immune cells secretion of DNA networks, as we demonstrate by trapping E. coli bacteria., Lipid membrane disruption is often associated with disease but is also essential to a range of biosensing and therapeutic techniques. Here, the authors report on the development of DNA-based particles that, upon exposure to an external cue, can aggregate, disrupt lipid membranes, and arrest the motion of bacteria.
- Published
- 2021
5. Repeat DNA-PAINT suppresses background and non-specific signals in optical nanoscopy
- Author
-
William T. Kaufhold, Anna Meletiou, Alexander H. Clowsley, Lorenzo Di Michele, Tobias Lutz, Christian Soeller, Clowsley, Alexander H [0000-0001-9318-1557], Meletiou, Anna [0000-0003-1167-0555], Di Michele, Lorenzo [0000-0002-1458-9747], Soeller, Christian [0000-0002-9302-2203], Apollo - University of Cambridge Repository, Commission of the European Communities, and The Royal Society
- Subjects
0301 basic medicine ,Oligonucleotides ,General Physics and Astronomy ,02 engineering and technology ,01 natural sciences ,Fluorescence imaging ,Reduction (complexity) ,chemistry.chemical_compound ,Non specific ,Nucleic Acids ,Nanotechnology ,Super-resolution microscopy ,Image resolution ,Physics ,0303 health sciences ,Multidisciplinary ,Sampling (statistics) ,021001 nanoscience & nanotechnology ,Fluorescence ,humanities ,3. Good health ,Multidisciplinary Sciences ,Science & Technology - Other Topics ,0210 nano-technology ,Biological system ,SUPERRESOLUTION MICROSCOPY ,Biotechnology ,Materials science ,Science ,010402 general chemistry ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Animals ,Spurious relationship ,HYBRIDIZATION KINETICS ,030304 developmental biology ,Science & Technology ,Reproducibility of Results ,General Chemistry ,DNA ,0104 chemical sciences ,Nanostructures ,030104 developmental biology ,RESOLUTION ,chemistry ,Microscopy, Fluorescence ,Biophysics ,Transient (oscillation) - Abstract
DNA-PAINT is a versatile optical super-resolution technique relying on the transient binding of fluorescent DNA ‘imagers’ to target epitopes. Its performance in biological samples is often constrained by strong background signals and non-specific binding events, both exacerbated by high imager concentrations. Here we describe Repeat DNA-PAINT, a method that enables a substantial reduction in imager concentration, thus suppressing spurious signals. Additionally, Repeat DNA-PAINT reduces photoinduced target-site loss and can accelerate sampling, all without affecting spatial resolution., DNA-PAINT is a super-resolution imaging technique which suffers from high background signals and non-specific binding. Here the authors report Repeat DNA-PAINT which is capable of supressing background noise and preventing photoinduced site loss, as well as decreasing the time taken for the sampling process.
- Published
- 2020
- Full Text
- View/download PDF
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