Kinetic Analysis and Intermediate Structure Determination From High-Speed Time-Resolved Crystallography
Abstract and Figures
Despite the very recent availability of High Speed Time-Resolved Crystallography (HSTC) data sets, the promise of HSTC remains largely unfulfilled due to the difficulty in extracting structural information on intermediates from time-resolved electron density maps. These maps are only composites of the electron density maps of the populated chemical species present at the experimental time points. Although various schemes of extracting homogeneous structures from HSTC data sets have been proposed, to date no structures have been determined. In this paper we present novel methods for estimating these parameters directly from an HSTC data set. We apply our methods to successfully extract structural information on intermediates from an HSTC data set acquired during decay from the saturated photostationary state of the Photoactive Yellow Protein (PYP). We present the refined structure (pseudo-2.0 Angstrom resolution by estimates of completeness) for a previously unseen intermediate of the PYP photocycle decay.
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