Structural enzymology using X-ray free electron lasers

Christopher Kupitz, Jose L. Olmos, Mark Holl, Lee Tremblay, Kanupriya Pande, Suraj Pandey, Dominik Oberthür, Mark Hunter, Mengning Liang, Andrew Aquila, Jason Tenboer, George Calvey, Andrea Katz, Yujie Chen, Max O. Wiedorn, Juraj Knoska, Alke Meents, Valerio Majriani, Tyler Norwood, Ishwor Poudyal & 30 others Thomas Grant, Mitchell D. Miller, Weijun Xu, Aleksandra Tolstikova, Andrew Morgan, Markus Metz, Jose Martin-Gracia, James D. Zook, Shatabdi Roy-Chowdhury, Jesse Coe, Nirupa Nagaratnam, Domingo Meza, Raimund Fromme, Shibom Basu, Matthias Frank, Thomas White, Anton Barty, Sasa Bajt, Oleksandr Yefanov, Henry N. Chapman, Nadia Zatsepin, Garrett Nelson, Uwe Weierstall, John Spence, Peter Schwander, Lois Pollack, Petra Fromme, Abbas Ourmazd, George N. Phillips, Marius Schmidt

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Mix-and-inject serial crystallography (MISC) is a technique designed to image enzyme catalyzed reactions in which small protein crystals are mixed with a substrate just prior to being probed by an X-ray pulse. This approach offers several advantages over flow cell studies. It provides (i) room temperature structures at near atomic resolution, (ii) time resolution ranging from microseconds to seconds, and (iii) convenient reaction initiation. It outruns radiation damage by using femtosecond X-ray pulses allowing damage and chemistry to be separated. Here, we demonstrate that MISC is feasible at an X-ray free electron laser by studying the reaction of M. tuberculosis ß-lactamase microcrystals with ceftriaxone antibiotic solution. Electron density maps of the apo-ß-lactamase and of the ceftriaxone bound form were obtained at 2.8 Å and 2.4 Å resolution, respectively. These results pave the way to study cyclic and non-cyclic reactions and represent a new field of time-resolved structural dynamics for numerous substrate-triggered biological reactions.

Original languageEnglish (US)
Article number044003
JournalStructural Dynamics
Volume4
Issue number4
DOIs
StatePublished - Jul 1 2017

Fingerprint

enzymology
X ray lasers
Crystallography
Ceftriaxone
Free electron lasers
free electron lasers
X rays
Microcrystals
Radiation damage
Structural dynamics
Antibiotics
Substrates
Carrier concentration
x rays
Enzymes
crystallography
Anti-Bacterial Agents
Proteins
Crystals
tuberculosis

ASJC Scopus subject areas

  • Radiation
  • Instrumentation
  • Condensed Matter Physics
  • Spectroscopy

Cite this

Kupitz, C., Olmos, J. L., Holl, M., Tremblay, L., Pande, K., Pandey, S., ... Schmidt, M. (2017). Structural enzymology using X-ray free electron lasers. Structural Dynamics, 4(4), [044003]. https://doi.org/10.1063/1.4972069

Structural enzymology using X-ray free electron lasers. / Kupitz, Christopher; Olmos, Jose L.; Holl, Mark; Tremblay, Lee; Pande, Kanupriya; Pandey, Suraj; Oberthür, Dominik; Hunter, Mark; Liang, Mengning; Aquila, Andrew; Tenboer, Jason; Calvey, George; Katz, Andrea; Chen, Yujie; Wiedorn, Max O.; Knoska, Juraj; Meents, Alke; Majriani, Valerio; Norwood, Tyler; Poudyal, Ishwor; Grant, Thomas; Miller, Mitchell D.; Xu, Weijun; Tolstikova, Aleksandra; Morgan, Andrew; Metz, Markus; Martin-Gracia, Jose; Zook, James D.; Roy-Chowdhury, Shatabdi; Coe, Jesse; Nagaratnam, Nirupa; Meza, Domingo; Fromme, Raimund; Basu, Shibom; Frank, Matthias; White, Thomas; Barty, Anton; Bajt, Sasa; Yefanov, Oleksandr; Chapman, Henry N.; Zatsepin, Nadia; Nelson, Garrett; Weierstall, Uwe; Spence, John; Schwander, Peter; Pollack, Lois; Fromme, Petra; Ourmazd, Abbas; Phillips, George N.; Schmidt, Marius.

In: Structural Dynamics, Vol. 4, No. 4, 044003, 01.07.2017.

Research output: Contribution to journalArticle

Kupitz, C, Olmos, JL, Holl, M, Tremblay, L, Pande, K, Pandey, S, Oberthür, D, Hunter, M, Liang, M, Aquila, A, Tenboer, J, Calvey, G, Katz, A, Chen, Y, Wiedorn, MO, Knoska, J, Meents, A, Majriani, V, Norwood, T, Poudyal, I, Grant, T, Miller, MD, Xu, W, Tolstikova, A, Morgan, A, Metz, M, Martin-Gracia, J, Zook, JD, Roy-Chowdhury, S, Coe, J, Nagaratnam, N, Meza, D, Fromme, R, Basu, S, Frank, M, White, T, Barty, A, Bajt, S, Yefanov, O, Chapman, HN, Zatsepin, N, Nelson, G, Weierstall, U, Spence, J, Schwander, P, Pollack, L, Fromme, P, Ourmazd, A, Phillips, GN & Schmidt, M 2017, 'Structural enzymology using X-ray free electron lasers' Structural Dynamics, vol. 4, no. 4, 044003. https://doi.org/10.1063/1.4972069
Kupitz C, Olmos JL, Holl M, Tremblay L, Pande K, Pandey S et al. Structural enzymology using X-ray free electron lasers. Structural Dynamics. 2017 Jul 1;4(4). 044003. https://doi.org/10.1063/1.4972069
Kupitz, Christopher ; Olmos, Jose L. ; Holl, Mark ; Tremblay, Lee ; Pande, Kanupriya ; Pandey, Suraj ; Oberthür, Dominik ; Hunter, Mark ; Liang, Mengning ; Aquila, Andrew ; Tenboer, Jason ; Calvey, George ; Katz, Andrea ; Chen, Yujie ; Wiedorn, Max O. ; Knoska, Juraj ; Meents, Alke ; Majriani, Valerio ; Norwood, Tyler ; Poudyal, Ishwor ; Grant, Thomas ; Miller, Mitchell D. ; Xu, Weijun ; Tolstikova, Aleksandra ; Morgan, Andrew ; Metz, Markus ; Martin-Gracia, Jose ; Zook, James D. ; Roy-Chowdhury, Shatabdi ; Coe, Jesse ; Nagaratnam, Nirupa ; Meza, Domingo ; Fromme, Raimund ; Basu, Shibom ; Frank, Matthias ; White, Thomas ; Barty, Anton ; Bajt, Sasa ; Yefanov, Oleksandr ; Chapman, Henry N. ; Zatsepin, Nadia ; Nelson, Garrett ; Weierstall, Uwe ; Spence, John ; Schwander, Peter ; Pollack, Lois ; Fromme, Petra ; Ourmazd, Abbas ; Phillips, George N. ; Schmidt, Marius. / Structural enzymology using X-ray free electron lasers. In: Structural Dynamics. 2017 ; Vol. 4, No. 4.
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abstract = "Mix-and-inject serial crystallography (MISC) is a technique designed to image enzyme catalyzed reactions in which small protein crystals are mixed with a substrate just prior to being probed by an X-ray pulse. This approach offers several advantages over flow cell studies. It provides (i) room temperature structures at near atomic resolution, (ii) time resolution ranging from microseconds to seconds, and (iii) convenient reaction initiation. It outruns radiation damage by using femtosecond X-ray pulses allowing damage and chemistry to be separated. Here, we demonstrate that MISC is feasible at an X-ray free electron laser by studying the reaction of M. tuberculosis {\ss}-lactamase microcrystals with ceftriaxone antibiotic solution. Electron density maps of the apo-{\ss}-lactamase and of the ceftriaxone bound form were obtained at 2.8 {\AA} and 2.4 {\AA} resolution, respectively. These results pave the way to study cyclic and non-cyclic reactions and represent a new field of time-resolved structural dynamics for numerous substrate-triggered biological reactions.",
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