NIH support: P50GM073197

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The Joint Center for Innovative Membrane Protein Technologies-Complexes (JCIMPT-Complexes) is a research network comprising multiple integrated biophysical projects including single molecule spectroscopy, nuclear magnetic resonance, x-ray crystallography, small angle x-ray scattering, and electron microscopy all applied and optimized in the areas of membrane protein expression, stabilization, and biophysical characterization of membrane protein-soluble protein complexes.

The primary mission is to develop and disseminate novel and enabling methods and technologies to the scientific community that lead to the structure determination of human membrane proteins and their membrane protein-soluble protein complexes. Careful characterization, miniaturization and automation are the major themes in the systematic development of new technologies to study membrane proteins and membrane protein complexes.
Focus area A will work on production of eukaryotic (particularly human) monomeric, homomeric, and heteromeric membrane proteins and their complexes. A major effort will be in completing and reducing to a robust method the JCIMPT protocol of using parallel microexpression, characterization, and purification of human membrane protein constructs for selecting the best construct for producing structure grade protein; thus reducing cost and effort, and increasing success rates. Single molecule spectroscopic studies will be used to characterize and understand the nature of multi-protein complexes with the goal of developing technology to evaluate and produce functional and stable assemblies.
Focus area B will work on design and validation of new compounds (e.g. lipids, detergents, lipidic cubic phase) for stabilizing membrane proteins. New compounds that form lipid cubic phase, as well as sponge phase, will be designed and tested in stabilization and crystallization studies.
Focus area C will work on characterization and structure solution of membrane proteins and complexes using NMR spectroscopy, X-ray diffraction, small angle scattering, and electron microscopy. Finally, a new outreach program will disseminate knowledge (e.g. methods and protocols) as well as reagents. Over the five-year funding period, we will conduct workshops and host meetings, as well as maintain a comprehensive and up-to-date website that will become a powerful portal for those scientists interested in membrane protein structural biology.

Project activity summarizing the three major focus areas (A, B, C) and seven specific projects (1-7) for JCIMPT-Complexes. Single molecule analysis of complexes using fluoresence based methods (Project 2) is shown in both focus areas A and C given its role in helping to understand and select for constructs and conditions in vivo, during focus A expression development as well as the focus C in vitro characterization. To avoid duplication in the proposal, this project is listed as project 2 where it is initially presented.

The JCIMPT is supported by the National Institutes of Health, NIH Common Fund -- Roadmap for Structural Biology, grant number P50GM073197