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Heterogeneity in M. tuberculosis β-lactamase inhibition by Sulbactam

By Tek Narsingh Malla1, Kara Zielinski2, Luis Aldama, Sasa Bajt, Denisse Feliz, Brendon Hayes, Mark S. Hunter3, Christopher Kupitz3, Stella Lisova, Juraj Knoska, Jose Manuel Martin-Garcia, Valerio Mariani, Suraj Pandey1, Ishwor Poudyal1, Raymond G. Sierra, Alexandra Tolstikova, Oleksandr Yefanov, Chung Hong Yoon, Abbas Ourmazd4, Petra Fromme5, Peter Schwander4, Anton Barty, Henry Chapman2, Emina Stojkovic6, Alexander Batyuk, Sébastien Boutet, George Phillips7, Lois Pollack8, Marius Schmidt1

1. University of Wisconsin - Milwaukee 2. Center for Free-Electron Laser Science 3. SLAC National Accelerator Laboratory 4. University of Wisconsin-Milwaukee 5. Arizona State University 6. Northern Illinois University 7. Rice University 8. Cornell University

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Category

BioXFEL Publications

Published on

Type

journal-article

Author

Tek Narsingh Malla and Kara Zielinski and Luis Aldama and Sasa Bajt and Denisse Feliz and Brendon Hayes and Mark Hunter and Christopher Kupitz and Stella Lisova and Juraj Knoska and Jose Manuel Martin-Garcia and Valerio Mariani and Suraj Pandey and Ishwor Poudyal and Raymond G. Sierra and Alexandra Tolstikova and Oleksandr Yefanov and Chung Hong Yoon and Abbas Ourmazd and Petra Fromme and Peter Schwander and Anton Barty and Henry N. Chapman and Emina A. Stojkovic and Alexander Batyuk and Sébastien Boutet and George N. Phillips and Lois Pollack and Marius Schmidt

Citation

Malla, T. N., Zielinski, K., Aldama, L., Bajt, S., Feliz, D., Hayes, B., Hunter, M., Kupitz, C., Lisova, S., Knoska, J., Martin-Garcia, J. M., Mariani, V., Pandey, S., Poudyal, I., Sierra, R. G., Tolstikova, A., Yefanov, O., Yoon, C. H., Ourmazd, A., … Schmidt, M. (2023). Heterogeneity in M. tuberculosis β-lactamase inhibition by Sulbactam. Nature Communications, 14(1). https://doi.org/10.1038/s41467-023-41246-1

Abstract

AbstractFor decades, researchers have elucidated essential enzymatic functions on the atomic length scale by tracing atomic positions in real-time. Our work builds on possibilities unleashed by mix-and-inject serial crystallography (MISC) at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals. Here, we report in atomic detail (between 2.2 and 2.7 Å resolution) by room-temperature, time-resolved crystallography with millisecond time-resolution (with timepoints between 3 ms and 700 ms) how the Mycobacterium tuberculosis enzyme BlaC is inhibited by sulbactam (SUB). Our results reveal ligand binding heterogeneity, ligand gating, cooperativity, induced fit, and conformational selection all from the same set of MISC data, detailing how SUB approaches the catalytic clefts and binds to the enzyme noncovalently before reacting to a trans-enamine. This was made possible in part by the application of singular value decomposition to the MISC data using a program that remains functional even if unit cell parameters change up to 3 Å during the reaction.

DOI

http://dx.doi.org/10.1038/s41467-023-41246-1

Funding

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