Enzyme intermediates captured "on the fly" by mix-and-inject serial crystallography

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 & 30 others 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

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: 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. Results: 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. Conclusions: 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.

Original languageEnglish (US)
Article number59
JournalBMC Biology
Volume16
Issue number1
DOIs
StatePublished - May 31 2018
Externally publishedYes

Fingerprint

crystallography
Crystallography
Diptera
X-radiation
X-Rays
enzyme
antibiotics
Enzymes
enzymes
Anti-Bacterial Agents
X ray lasers
ceftriaxone
Crystal atomic structure
X rays
Reaction intermediates
tuberculosis
Ceftriaxone
Drug Design
Free electron lasers
Mycobacterium tuberculosis

ASJC Scopus subject areas

  • Biotechnology
  • Structural Biology
  • Ecology, Evolution, Behavior and Systematics
  • Physiology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Plant Science
  • Developmental Biology
  • Cell Biology

Cite this

Olmos, J. L., Pandey, S., Martin-Garcia, J. M., Calvey, G., Katz, A., Knoska, J., ... Schmidt, M. (2018). Enzyme intermediates captured "on the fly" by mix-and-inject serial crystallography. BMC Biology, 16(1), [59]. https://doi.org/10.1186/s12915-018-0524-5

Enzyme intermediates captured "on the fly" by mix-and-inject serial crystallography. / Olmos, Jose L.; Pandey, Suraj; Martin-Garcia, Jose M.; Calvey, George; Katz, Andrea; Knoska, Juraj; Kupitz, Christopher; Hunter, Mark S.; Liang, Mengning; Oberthuer, Dominik; Yefanov, Oleksandr; Wiedorn, Max; Heyman, Michael; Holl, Mark; Pande, Kanupriya; Barty, Anton; Miller, Mitchell D.; Stern, Stephan; Roy-Chowdhury, Shatabdi; Coe, Jesse; Nagaratnam, Nirupa; Zook, James; Verburgt, Jacob; Norwood, Tyler; Poudyal, Ishwor; Xu, David; Koglin, Jason; Seaberg, Matthew H.; Zhao, Yun; Bajt, Saša; Grant, Thomas; Mariani, Valerio; Nelson, Garrett; Subramanian, Ganesh; Bae, Euiyoung; Fromme, Raimund; Fung, Russell; Schwander, Peter; Frank, Matthias; White, Thomas A.; Weierstall, Uwe; Zatsepin, Nadia; Spence, John; Fromme, Petra; Chapman, Henry N.; Pollack, Lois; Tremblay, Lee; Ourmazd, Abbas; Phillips, George N.; Schmidt, Marius.

In: BMC Biology, Vol. 16, No. 1, 59, 31.05.2018.

Research output: Contribution to journalArticle

Olmos, JL, Pandey, S, Martin-Garcia, JM, Calvey, G, Katz, A, Knoska, J, Kupitz, C, Hunter, MS, Liang, M, Oberthuer, D, Yefanov, O, Wiedorn, M, Heyman, M, Holl, M, Pande, K, Barty, A, Miller, MD, Stern, S, Roy-Chowdhury, S, Coe, J, Nagaratnam, N, Zook, J, Verburgt, J, Norwood, T, Poudyal, I, Xu, D, Koglin, J, Seaberg, MH, Zhao, Y, Bajt, S, Grant, T, Mariani, V, Nelson, G, Subramanian, G, Bae, E, Fromme, R, Fung, R, Schwander, P, Frank, M, White, TA, Weierstall, U, Zatsepin, N, Spence, J, Fromme, P, Chapman, HN, Pollack, L, Tremblay, L, Ourmazd, A, Phillips, GN & Schmidt, M 2018, 'Enzyme intermediates captured "on the fly" by mix-and-inject serial crystallography', BMC Biology, vol. 16, no. 1, 59. https://doi.org/10.1186/s12915-018-0524-5
Olmos JL, Pandey S, Martin-Garcia JM, Calvey G, Katz A, Knoska J et al. Enzyme intermediates captured "on the fly" by mix-and-inject serial crystallography. BMC Biology. 2018 May 31;16(1). 59. https://doi.org/10.1186/s12915-018-0524-5
Olmos, Jose L. ; Pandey, Suraj ; Martin-Garcia, Jose M. ; Calvey, George ; Katz, Andrea ; Knoska, Juraj ; Kupitz, Christopher ; Hunter, Mark S. ; Liang, Mengning ; Oberthuer, Dominik ; Yefanov, Oleksandr ; Wiedorn, Max ; Heyman, Michael ; Holl, Mark ; Pande, Kanupriya ; Barty, Anton ; Miller, Mitchell D. ; Stern, Stephan ; Roy-Chowdhury, Shatabdi ; Coe, Jesse ; Nagaratnam, Nirupa ; Zook, James ; Verburgt, Jacob ; Norwood, Tyler ; Poudyal, Ishwor ; Xu, David ; Koglin, Jason ; Seaberg, Matthew H. ; Zhao, Yun ; Bajt, Saša ; Grant, Thomas ; Mariani, Valerio ; Nelson, Garrett ; Subramanian, Ganesh ; Bae, Euiyoung ; Fromme, Raimund ; Fung, Russell ; Schwander, Peter ; Frank, Matthias ; White, Thomas A. ; Weierstall, Uwe ; Zatsepin, Nadia ; Spence, John ; Fromme, Petra ; Chapman, Henry N. ; Pollack, Lois ; Tremblay, Lee ; Ourmazd, Abbas ; Phillips, George N. ; Schmidt, Marius. / Enzyme intermediates captured "on the fly" by mix-and-inject serial crystallography. In: BMC Biology. 2018 ; Vol. 16, No. 1.
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AU - Pandey, Suraj

AU - Martin-Garcia, Jose M.

AU - Calvey, George

AU - Katz, Andrea

AU - Knoska, Juraj

AU - Kupitz, Christopher

AU - Hunter, Mark S.

AU - Liang, Mengning

AU - Oberthuer, Dominik

AU - Yefanov, Oleksandr

AU - Wiedorn, Max

AU - Heyman, Michael

AU - Holl, Mark

AU - Pande, Kanupriya

AU - Barty, Anton

AU - Miller, Mitchell D.

AU - Stern, Stephan

AU - Roy-Chowdhury, Shatabdi

AU - Coe, Jesse

AU - Nagaratnam, Nirupa

AU - Zook, James

AU - Verburgt, Jacob

AU - Norwood, Tyler

AU - Poudyal, Ishwor

AU - Xu, David

AU - Koglin, Jason

AU - Seaberg, Matthew H.

AU - Zhao, Yun

AU - Bajt, Saša

AU - Grant, Thomas

AU - Mariani, Valerio

AU - Nelson, Garrett

AU - Subramanian, Ganesh

AU - Bae, Euiyoung

AU - Fromme, Raimund

AU - Fung, Russell

AU - Schwander, Peter

AU - Frank, Matthias

AU - White, Thomas A.

AU - Weierstall, Uwe

AU - Zatsepin, Nadia

AU - Spence, John

AU - Fromme, Petra

AU - Chapman, Henry N.

AU - Pollack, Lois

AU - Tremblay, Lee

AU - Ourmazd, Abbas

AU - Phillips, George N.

AU - Schmidt, Marius

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