Title |
Enzyme intermediates captured “on the fly” by mix-and-inject serial crystallography
|
---|---|
Published in |
BMC Biology, May 2018
|
DOI | 10.1186/s12915-018-0524-5 |
Pubmed ID | |
Authors |
Jose L. Olmos, Suraj Pandey, Jose M. Martin-Garcia, George Calvey, Andrea Katz, Juraj Knoska, Christopher Kupitz, Mark S. Hunter, Mengning Liang, Dominik Oberthuer, Oleksandr Yefanov, Max Wiedorn, Michael Heyman, Mark Holl, Kanupriya Pande, Anton Barty, Mitchell D. Miller, Stephan Stern, Shatabdi Roy-Chowdhury, Jesse Coe, Nirupa Nagaratnam, James Zook, Jacob Verburgt, Tyler Norwood, Ishwor Poudyal, David Xu, Jason Koglin, Matthew H. Seaberg, Yun Zhao, Saša Bajt, Thomas Grant, Valerio Mariani, Garrett Nelson, Ganesh Subramanian, Euiyoung Bae, Raimund Fromme, Russell Fung, Peter Schwander, Matthias Frank, Thomas A. White, Uwe Weierstall, Nadia Zatsepin, John Spence, Petra Fromme, Henry N. Chapman, Lois Pollack, Lee Tremblay, Abbas Ourmazd, George N. Phillips, Marius Schmidt |
Abstract |
Ever since the first atomic structure of an enzyme was solved, the discovery of the mechanism and dynamics of reactions catalyzed by biomolecules has been the key goal for the understanding of the molecular processes that drive life on earth. Despite a large number of successful methods for trapping reaction intermediates, the direct observation of an ongoing reaction has been possible only in rare and exceptional cases. Here, we demonstrate a general method for capturing enzyme catalysis "in action" by mix-and-inject serial crystallography (MISC). Specifically, we follow the catalytic reaction of the Mycobacterium tuberculosis β-lactamase with the third-generation antibiotic ceftriaxone by time-resolved serial femtosecond crystallography. The results reveal, in near atomic detail, antibiotic cleavage and inactivation from 30 ms to 2 s. MISC is a versatile and generally applicable method to investigate reactions of biological macromolecules, some of which are of immense biological significance and might be, in addition, important targets for structure-based drug design. With megahertz X-ray pulse rates expected at the Linac Coherent Light Source II and the European X-ray free-electron laser, multiple, finely spaced time delays can be collected rapidly, allowing a comprehensive description of biomolecular reactions in terms of structure and kinetics from the same set of X-ray data. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 3 | 20% |
United Kingdom | 3 | 20% |
South Africa | 1 | 7% |
Indonesia | 1 | 7% |
Germany | 1 | 7% |
Unknown | 6 | 40% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 11 | 73% |
Scientists | 2 | 13% |
Science communicators (journalists, bloggers, editors) | 2 | 13% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 137 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 37 | 27% |
Researcher | 31 | 23% |
Student > Master | 13 | 9% |
Student > Doctoral Student | 7 | 5% |
Professor | 6 | 4% |
Other | 11 | 8% |
Unknown | 32 | 23% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 47 | 34% |
Physics and Astronomy | 17 | 12% |
Agricultural and Biological Sciences | 16 | 12% |
Chemistry | 10 | 7% |
Materials Science | 3 | 2% |
Other | 12 | 9% |
Unknown | 32 | 23% |