Time-resolved serial crystallography as a tool for mechanistic enzymology
by
Auditorium, Lighthouse
European XFEL
Enzyme catalysis is essential for life and is a central phenomenon in biochemistry. The advent of time-resolved serial crystallography, initially enabled by X-ray free electron lasers (XFELs) and now expanding to synchrotron X-ray sources, allows enzyme catalysis to be observed in real time, in near-physiological conditions, and at atomic resolution. I will describe our work using mix-and-inject serial crystallography (MISC) to observe catalysis by two proteins from the DJ-1 superfamily: isocyanide hydratase (ICH) and human DJ-1. In ICH, MISC allowed us to observe formation of a hypothetical catalytic intermediate and to determine how enzyme conformational strain interacts with active site electrostatic changes to gate non-equilibrium conformational dynamics during catalysis. In human DJ-1, MISC at a synchrotron permitted the observation of catalytic intermediates that resolve a long-standing mechanistic controversy about the protein. Synchrotron deployment of MISC with DJ-1 illustrates both the promise of broad access to this important technology as well as some considerations for radiation-sensitive samples. Time-resolved X-ray crystallography is entering an era of wider accessibility and has the potential to make non-equilibrium studies a major growth area for the next phase of structural biology.
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