Observation of substrate diffusion and ligand binding in enzyme crystals using high-repetition-rate mix-and-inject serial crystallography

Pandey, S.; Calvey, G.; Katz, A.M.; Malla, T.N.; Koua, F.H.M.; Martin-Garcia, J.M.; Poudyal, I.; Yang, J.-H.; Vakili, M.; Yefanov, O.; Zielinski, K.A.; Bajt, S.; Awel, S.; Doerner, K.; Frank, M.; Gelisio, L.; Jernigan, R.; Kirkwood, H.; Kloos, M.; Koliyadu, J.; Mariani, V.; Miller, M.D.; Mills, G.; Nelson, G.; Olmos Jr., J.L.; Sadri, A.; Sato, T.; Tolstikova, A.; Xu, W.; Ourmazd, A.; Spence, J.H.C.; Schwander, P.; Barty, A.; Chapman, H.N.; Fromme, P.; Mancuso, A.P.; Phillips Jr, G.N.; Bean, R.; Pollack, L.; Schmidt, M.

doi:10.1107/S2052252521008125

Figure 3
Simplified enzymatic cycle of, and the timescales associated with, the reaction of BlaC with CEF. CEF is delivered to the crystals by diffusion. It noncovalently binds to the free BlaC enzyme (E) to form the enzyme–substrate complex (ES). The acyl intermediate EP, which is covalently bound to Ser70, is formed within ∼200 ms. The leaving group R₂ is cleaved off the CEF. The modified CEF (EP) is hydrolyzed and released as product (P) and the free enzyme is recovered within about 2 s. In this paper, only the formation of the ES complex up to 50 ms (gray, blurred vertical line) was explored.

IUCrJ

Volume 8| Part 6| November 2021| Pages 878-895

ISSN: 2052-2525

https://doi.org/10.1107/S2052252521008125

BIOLOGY | MEDICINE

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