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

PloS One
Mikhail SergeevIra Kurtz

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.

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Citations

Feb 12, 2014·Langmuir : the ACS Journal of Surfaces and Colloids·Katarzyna ZielińskaHerman P van Leeuwen
Aug 8, 2013·Current Opinion in Nephrology and Hypertension·Alan S L Yu
Aug 7, 2013·Current Opinion in Nephrology and Hypertension·Ira Kurtz, Quansheng Zhu
Apr 16, 2013·BioTechniques·Cesar TrigoSebastian Brauchi
Mar 4, 2018·Nature Communications·Kevin W HuynhIra Kurtz
Apr 17, 2013·Physiological Reviews·Mark D Parker, Walter F Boron

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Methods Mentioned

BETA
FRET
fluorescence correlation spectroscopy
FCS
fluorescence microscopy
glycosylation
transfection
transfections

Software Mentioned

SpIDA
MATLAB

Related Concepts

Related Papers

Biophysical Journal
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Proceedings of the National Academy of Sciences of the United States of America
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Proceedings of the National Academy of Sciences of the United States of America
Jody L SwiftJean-Martin Beaulieu
Proceedings of the National Academy of Sciences of the United States of America
Kyoko ShirakabeKatsuhiko Mikoshiba
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