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PLoS One. 2012;7(4):e36215. doi: 10.1371/journal.pone.0036215. Epub 2012 Apr 27.

Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis.

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1
Department of Physics, McGill University, Montréal, Québec, Canada.

Abstract

Membrane transporter proteins exist in a complex dynamic equilibrium between various oligomeric states that include monomers, dimers, dimer of dimers and higher order oligomers. Given their sub-optical microscopic resolution size, the oligomerization state of membrane transporters is difficult to quantify without requiring tissue disruption and indirect biochemical methods. Here we present the application of a fluorescence measurement technique which combines fluorescence image moment analysis and spatial intensity distribution analysis (SpIDA) to determine the oligomerization state of membrane proteins in situ. As a model system we analyzed the oligomeric state(s) of the electrogenic sodium bicarbonate cotransporter NBCe1-A in cultured cells and in rat kidney. The approaches that we describe offer for the first time the ability to investigate the oligomeric state of membrane transporter proteins in their native state.

PMID:
22558387
PMCID:
PMC3338697
DOI:
10.1371/journal.pone.0036215
[Indexed for MEDLINE]
Free PMC Article
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