Measuring Membrane Protein Dimerization Equilibrium in Lipid Bilayers by Single-Molecule Fluorescence Microscopy

Content Provider	Scilit
Author	Chadda, R. Robertson, J. L.
Copyright Year	2016
Description	Dimerization of membrane protein interfaces occurs during membrane protein folding and cell receptor signaling. Here, we summarize a method that allows for measurement of equilibrium dimerization reactions of membrane proteins in lipid bilayers, by measuring the Poisson distribution of subunit capture into liposomes by single-molecule photobleaching analysis. This strategy is grounded in the fact that given a comparable labeling efficiency, monomeric or dimeric forms of a membrane protein will give rise to distinctly different photobleaching probability distributions. These methods have been used to verify the dimer stoichiometry of the Fluc $F^{−}$ ion channel and the dimerization equilibrium constant of the ClC-ec1 $Cl^{−}/H^{+}$ antiporter in lipid bilayers. This approach can be applied to any membrane protein system provided it can be purified, fluorescently labeled in a quantitative manner, and verified to be correctly folded by functional assays, even if the structure is not yet known. Book Name: Protein \3- DNA Interactions
Related Links	http://europepmc.org/articles/pmc5568537?pdf=render https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5568537/pdf
Ending Page	82
Page Count	30
Starting Page	53
DOI	10.1016/bs.mie.2016.08.025
Volume Number	581
Language	English
Publisher	Elsevier BV
Publisher Date	2016-10-11
Access Restriction	Open
Subject Keyword	Book Name: Protein \3- DNA Interactions Single-molecule Microscopy
Content Type	Text
Resource Type	Article