Electron crystallography has a key role to play in understanding integral membrane proteins (IMPs) as it provides one of the most direct means of providing insight into the functional state of these molecular machines in their lipid-associated forms, and also has the potential to facilitate examination of physiologically relevant transitional states and complexes. Helical or tubular crystals, which are the natural product of proteins crystallizing on the surface of a vesicle, offer some unique advantages over other crystalline forms. The goal of this proposal is to develop an integrated pipeline to enable structure determination by transmission electron microcopy of IMPs in the form of tubular crystals. Given the very high number of IMPs that need to be solved, helical crystallization should be available as a standard method in the armamentarium of approaches for determining the structure of IMPs. However, its use is currently limited by a lack of standardized approaches and the enormous effort and specialized expertise required that form a barrier to entry. New or streamlined approaches are needed in three areas: crystal growth, screening, and analysis. An integrated approach is essential to ensure that progress in any one area is not throttled by bottlenecks at significant other steps in the process. Our goal is to establish TEM of tubular crystals as a standard approach to complement X-ray crystallography and NMR for characterizing IMPs.

Public Health Relevance

Membrane proteins account for ~30% of gene products in organisms and are the target of ~60% of all therapeutic drugs currently in development. This research will establish helical crystallization as a standard method to add to the armamentarium of approaches for determining the structure of these critically important macromolecular machines.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Special Emphasis Panel (ZRG1-BCMB-S (50))
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Chin, Jean
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Scripps Research Institute
La Jolla
United States
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Garnham, Christopher P; Vemu, Annapurna; Wilson-Kubalek, Elizabeth M et al. (2015) Multivalent Microtubule Recognition by Tubulin Tyrosine Ligase-like Family Glutamylases. Cell 161:1112-1123
Mulligan, Sean K; Speir, Jeffrey A; Razinkov, Ivan et al. (2015) Multiplexed TEM Specimen Preparation and Analysis of Plasmonic Nanoparticles. Microsc Microanal 21:1017-1025
Moeller, Arne; Lee, Sung Chang; Tao, Houchao et al. (2015) Distinct conformational spectrum of homologous multidrug ABC transporters. Structure 23:450-60
Lyumkis, Dmitry; Talley, Heather; Stewart, Andrew et al. (2013) Allosteric regulation of DNA cleavage and sequence-specificity through run-on oligomerization. Structure 21:1848-58
Tao, Houchao; Lee, Sung Chang; Moeller, Arne et al. (2013) Engineered nanostructured ?-sheet peptides protect membrane proteins. Nat Methods 10:759-61
Moeller, Arne; Zhao, Chunxia; Fried, Michael G et al. (2012) Nucleotide-dependent conformational changes in the N-Ethylmaleimide Sensitive Factor (NSF) and their potential role in SNARE complex disassembly. J Struct Biol 177:335-43
Moeller, Arne; Kirchdoerfer, Robert N; Potter, Clinton S et al. (2012) Organization of the influenza virus replication machinery. Science 338:1631-4
Fisher, Lauren S; Ward, Andrew; Milligan, Ronald A et al. (2011) A helical processing pipeline for EM structure determination of membrane proteins. Methods 55:350-62