Dresden 2020 – scientific programme

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BP: Fachverband Biologische Physik

BP 33: Protein Structure and Dynamics

BP 33.10: Talk

Thursday, March 19, 2020, 12:30–12:45, ZEU 250

Watching an enzyme at work: Time-Resolved Serial Crystallography reveals water mediated allosteric regulation — •Henrike Müller-Werkmeister — Uni Potsdam, Institut für Chemie, Physikalische Chemie, Karl-Liebknecht-Str. 24-25, 14476 Potsdam

We have studied the homodimeric enzyme fluoroacetate dehalogenase by time-resolved serial synchrotron crystallography (TR-SSX). Using a fixed target based sample delivery [1] with an efficient interlacing pattern allowed us to realize "hit-and-return" (HARE) TR-SSX to cover the full timescale from 30 milliseconds to 30 seconds [2]. With a photocaged substrate for reaction initiation, four catalytic turnovers could be resolved [3]. The total of 18 independent structures not only provide unprecedented insight into the reaction mechanism, showing the substrate binding, the Michaelis-Menten-complex and the covalent intermediate, but also reveal the allosteric mechanism leading to half-the-sites reactivity. In fact, a molecular water wire can be observed that together with molecular breathing is clocked to the enzymatic reaction.

[1] I. Martiel, H. M. Müller-Werkmeister, A. E. Cohen, Acta Cryst. D, 2019, D75, 160*177 [2] E. C. Schulz*, P. Mehrabi*, H. M. Müller-Werkmeister*, F. Tellkamp, A. Jha, W. Stuart, E. Persch, R. De Gasparo, F. Diederich, E. F. Pai, R. J. D. Miller, Nature Methods, 2018, 15 (11), 901-904 [3] P. Mehrabi*, E. C. Schulz*, R. Dsouza, H. M. Müller-Werkmeister, F. Tellkamp, R. J. D. Miller, E. F. Pai, Science, 2019, 365 (6458), 1167-1170