The Joint Center for Integral Membrane Protein Technologies-Complexes (JCIMPT-Complexes) is a research network comprising seven integrated projects working in three focused areas of membrane protein expression, stabilization, and biophysical characterization. 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 complexes. Miniaturization and automation are the major themes in the systematic development of new technologies to study membrane proteins and membrane protein complexes. Our extensive experience with human G protein-coupled receptors and their complexes make them the ideal target protein family for the planned focused technology development. In several active collaborations with others, we will work in parallel on targets including transporters and ion channels. 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.
Membrane proteins are the most difficult family of biological macromolecules to characterize at any level given their complex interaction with both the lipid and water soluble environments. These proteins (e.g. human G-protein coupled receptors) are the target of the majority of therapeutic drug targets. There is a critical need to develop breakthrough technologies to enable better characterization of this protein family.
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