A carboxy-terminal affinity tag for the purification and mass spectrometric characterization of integral membrane proteins.

ABSTRACT: G-protein-coupled receptors (GPCRs) and other structurally and functionally related membrane proteins represent particularly attractive targets for drug discovery. Integral membrane proteins are often difficult to purify from native contexts, and lack of sufficient quantities hampers subsequent structural and functional proteomic studies. We describe here an optimized enrichment strategy involving a membrane protein-compatible 1D4 affinity tag that is derived from the carboxy-terminal nine amino residues of bovine rhodopsin, and its corresponding tag-specific, high-affinity monoclonal antibody. When two GPCRs as well as two related ATP binding cassette (ABC) transporters are expressed in their functional forms in human cell lines, we have shown that a single detergent and wash condition ca

SUBMITTER: Wong JP 

PROVIDER: S-EPMC4103948 | biostudies-literature | 2009 May

REPOSITORIES: biostudies-literature