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Subunit counting in membrane-bound proteins.

ABSTRACT: The subunit number and stoichiometry of membrane-bound proteins are difficult to determine without disrupting their membrane environment. Here we describe a single-molecule technique for counting subunits of proteins in live cell membranes by observing bleaching steps of GFP fused to a protein of interest. After testing the method with proteins of known stoichiometry expressed in Xenopus laevis oocytes, we resolved the composition of NMDA receptors composed of NR1 and NR3 subunits.

SUBMITTER: Ulbrich MH 

PROVIDER: S-EPMC2744285 | biostudies-literature | 2007 Apr

REPOSITORIES: biostudies-literature

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Subunit counting in membrane-bound proteins.

Ulbrich Maximilian H MH   Isacoff Ehud Y EY  

Nature methods 20070318 4

The subunit number and stoichiometry of membrane-bound proteins are difficult to determine without disrupting their membrane environment. Here we describe a single-molecule technique for counting subunits of proteins in live cell membranes by observing bleaching steps of GFP fused to a protein of interest. After testing the method with proteins of known stoichiometry expressed in Xenopus laevis oocytes, we resolved the composition of NMDA receptors composed of NR1 and NR3 subunits. ...[more]

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