0 Citation(s)

0 questions (Ask a question)

Direct Observation of the Mechanism of Antibiotic Resistance by Mix-and-Inject at the European XFEL

By Suraj Pandey1, George Douglas Calvey, Andrea Katz, Tek Narsingh Malla1, Faisal Hammad Mekky Koua2, Jose M. Martin-Garcia, Ishwor Poudyal1, Jay-How Yang3, Mohammad Vakili, Oleksandr Yefanov, Kara Zielinski4, Saša Bajt, Salah Awel, Katerina Dörner, Matthias Frank5, Luca Gelisio, Rebecca Jeanne Jernigan3, Henry Kirkwood, Marco Kloos, Jayanath Koliyadu, Valerio Mariani, Mitchell Miller6, Grant Mills, Garrett Charles Nelson3, Jose L. Olmos, Alireza Sadri, Tokushi Sato, Alexandra Tolstikova, Weijun Xu, Abbas Ourmazd7, John Spence3, Peter Schwander7, Anton Barty, Henry Chapman4, Petra Fromme3, Adrian P. Mancuso, George Phillips6, Richard Bean, Lois Pollack8, Marius Schmidt1

1. University of Wisconsin - Milwaukee 2. Nagoya Institute of Technology 3. Arizona State University 4. Center for Free-Electron Laser Science 5. Lawrence Livermore National Laboratory 6. Rice University 7. University of Wisconsin-Milwaukee 8. Cornell University

See also

No results found.

Category

BioXFEL Publications

Published on

Type

posted-content

Author

Suraj Pandey and George Calvey and Andrea M. Katz and Tek Narsingh Malla and Faisal H. M. Koua and Jose M. Martin-Garcia and Ishwor Poudyal and Jay-How Yang and Mohammad Vakili and Oleksandr Yefanov and Kara A. Zielinski and Saša Bajt and Salah Awel and Katerina Dörner and Matthias Frank and Luca Gelisio and Rebecca Jernigan and Henry Kirkwood and Marco Kloos and Jayanath Koliyadu and Valerio Mariani and Mitchell D. Miller and Grant Mills and Garrett Nelson and Jose L. Olmos and Alireza Sadri and Tokushi Sato and Alexandra Tolstikova and Weijun Xu and Abbas Ourmazd and John H. C. Spence and Peter Schwander and Anton Barty and Henry N. Chapman and Petra Fromme and Adrian P. Mancuso and George N. Phillips and Richard Bean and Lois Pollack and Marius Schmidt

Citation

Pandey, S. et al., 2020. Direct Observation of the Mechanism of Antibiotic Resistance by Mix-and-Inject at the European XFEL. Available at: http://dx.doi.org/10.1101/2020.11.24.396689.

Abstract

AbstractIn this study, we follow the diffusion and buildup of occupancy of the substrate ceftriaxone in M. tuberculosis β-lactamase BlaC microcrystals by structural analysis of the enzyme substrate complex at single millisecond time resolution. We also show the binding and the reaction of an inhibitor, sulbactam, on a slower millisecond time scale. We use the ‘mix-and-inject’ technique to initiate these reactions by diffusion, and determine the resulting structures by serial crystallography using ultrafast, intense X-ray pulses from the European XFEL (EuXFEL) arriving at MHz repetition rates. Here, we show how to use the EuXFEL pulse structure to dramatically increase the size of the data set and thereby the quality and time resolution of “molecular movies” which unravel ligand binding and enzymatically catalyzed reactions. This shows the great potential for the EuXFEL as a tool for biomedically relevant research, particularly, as shown here, for investigating bacterial antibiotic resistance.One Sentence SummaryDirect observation of fast ligand binding in a biomedically relevant enzyme at near atomic resolution with MHz X-ray pulses at the European XFEL.

DOI

http://dx.doi.org/10.1101/2020.11.24.396689

Funding

NSF-STC Biology with X-ray Lasers (NSF-1231306)