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For decades, researchers have elucidated essential enzymatic functions on the atomic length scale by tracing atomic positions in real-time. Our work builds on possibilities unleashed by mix-and-inject serial crystallography (MISC) at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals. Here, we report in atomic detail (between 2.2 and 2.7 Å resolution) by room-temperature, time-resolved crystallography with millisecond time-resolution (with timepoints between 3 ms and 700 ms) how theMycobacterium tuberculosisenzyme BlaC is inhibited by sulbactam (SUB). Our results reveal ligand binding heterogeneity, ligand gating, cooperativity, induced fit, and conformational selection all from the same set of MISC data, detailing how SUB approaches the catalytic clefts and binds to the enzyme noncovalently before reacting to atrans-enamine. This was made possible in part by the application of singular value decomposition to the MISC data using a program that remains functional even if unit cell parameters change up to 3 Å during the reaction.

 

A 3D-printed modular droplet injector successfully delivered microcrystals of human NAD(P)H:quinone oxidoreductase 1 (NQO1) and phycocyanin with electrical stimulation in a serial crystallography experiment at 120 Hz repetition rate.

 

Let \(\Sigma\) be a closed subset of \(\mathbb{R}^{n+1}\) which is parabolic Ahlfors-David regular and assume that \(\Sigma\) satisfies a 2-sided corkscrew condition. Assume, in addition, that \(\Sigma\) is either time-forwards Ahlfors-David regular, time-backwards Ahlfors-David regular, or parabolic uniform rectifiable. We then first prove that \(\Sigma\) satisfies a weak synchronized two cube condition. Based on this we are able to revisit the argument of Nyström and Strömqvist (2009) and prove that \(\Sigma\) contain suniform big pieces of Lip(1,1/2) graphs. When \(\Sigma\) is parabolic uniformly rectifiable the construction can be refined and in this case we prove that \(\Sigma\) contains uniform big pieces of regular parabolic Lip(1,1/2) graphs. Similar results hold if \(\Omega\subset\mathbb{R}^{n+1}\) is a connected component of \(\mathbb{R}^{n+1}\setminus\Sigma\) and in this context we also give a parabolic counterpart of the main result of Azzam et al. (2017) by proving that if \(\Omega\) is a one-sided parabolic chord arc domain, and if \(\Sigma\) is parabolic uniformly rectifiable, then \(\Omega\) is in fact a parabolic chord arc domain. Our results give a flexible parabolic version of the classical (elliptic) result of David and Jerison (1990) concerning the existence of uniform big pieces of Lipschitz graphs for sets satisfying a two disc condition.