The overall objective of the proposed research is to develop enhanced anomalous diffraction methods for analyzing structures of biological macromolecules. We build on our recent accomplishments at New York Structural Biology Center (NYSBC) beamlines at Brookhaven's National Synchrotron Light Source (NSLS) and worldwide experience showing that multiwavelength anomalous diffraction (MAD) and SAD, MAD's single-wavelength counterpart, now predominate for de novo determinations of three- dimensional structures for biological macromolecules. We propose to optimize SAD and MAD phasing procedures by meeting the demands of compelling applications to current problems of biological significance. Biologically exciting problems motivate the development of appropriate tools, and forefront methods accelerate the solution of structures for systems of biological and medical significance. The overall objective is embodied in four specific aims: (1) we propose to enhance SAD phasing procedures for challenging problems such as selenomethionyl proteins at low resolution and only-light-atom native structures. A focus is on improved methods for increasing signal-to-noise ratios for anomalous diffraction by combining data from many crystals. (2) We propose to enhance MAD procedures for accurate experimental phase evaluation. Data collection strategies will be devised to mitigate radiation damage and minimize systematic errors. (3) We propose to develop procedures for low-energy anomalous diffraction experiments. We will develop experimental procedures and design instrumentation for low-energy (3 - 7 keV) experiments aiming to enhance anomalous signal from light atoms such as sulfur and phosphorous. (4) We propose to develop automation and enhanced procedures to facilitate SAD and MAD analyses from many crystals. A first priority is to encode and disseminate our current multi-crystal SAD process in convenient software.

Public Health Relevance

This is a project in technology development for basic science, but applications of expected to result from these studies are likely to have profound relevance for human health. Biological structures to be determined in atomic detail include molecular chaperones with impact for neurodegenerative diseases and cancer, calcium-release channel receptors with impact for heart failure, and histidine kinase receptors with relevance for microbial infection.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Macromolecular Structure and Function C Study Section (MSFC)
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Edmonds, Charles G
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Columbia University
Schools of Medicine
New York
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Su, Min; Gao, Feng; Yuan, Qi et al. (2017) Structural basis for conductance through TRIC cation channels. Nat Commun 8:15103
Liu, Qun; Hendrickson, Wayne A (2017) Contemporary Use of Anomalous Diffraction in Biomolecular Structure Analysis. Methods Mol Biol 1607:377-399
Liu, Qun (2017) TMBIM-mediated Ca2+ homeostasis and cell death. Biochim Biophys Acta 1864:850-857
Hendrickson, Wayne A (2016) Atomic-level analysis of membrane-protein structure. Nat Struct Mol Biol 23:464-7
Liu, Qun; Hendrickson, Wayne A (2015) Crystallographic phasing from weak anomalous signals. Curr Opin Struct Biol 34:99-107
Ranaivoson, Fanomezana M; Liu, Qun; Martini, Francesca et al. (2015) Structural and Mechanistic Insights into the Latrophilin3-FLRT3 Complex that Mediates Glutamatergic Synapse Development. Structure 23:1665-77
Guo, Youzhong; Kalathur, Ravi C; Liu, Qun et al. (2015) Protein structure. Structure and activity of tryptophan-rich TSPO proteins. Science 347:551-5
Yang, Tingting; Liu, Qun; Kloss, Brian et al. (2014) Structure and selectivity in bestrophin ion channels. Science 346:355-9
Liu, Qun; Guo, Youzhong; Chang, Yanqi et al. (2014) Multi-crystal native SAD analysis at 6?keV. Acta Crystallogr D Biol Crystallogr 70:2544-57
Zhang, Zhen; Liu, Qun; Hendrickson, Wayne A (2014) Crystal structures of apparent saccharide sensors from histidine kinase receptors prevalent in a human gut symbiont. FEBS J 281:4263-79

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