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PROTEOMIC METHOD FOR IDENTIFYING MEMBRANE PROTEINS

$10,719P41FY2009RRNIH

University Of Colorado, Boulder CO

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Abstract

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Integral membrane proteins perform crucial cellular functions and are the targets for the majority of pharmaceutical agents. However, the hydrophobic nature of their membrane-embedded domains makes them difficult to work with. Here, we describe a shotgun proteomic method for the high-throughput analysis of the membrane-embedded transmembrane domains of integral membrane proteins which extends the depth of coverage of the membrane proteome. Electron tomography and 3D reconstruction is used to deduce the 3D structure of the membrane preparations at different stages of the procedures. This work is currently in press in the Journal of Proteome Research.

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