Membrane Protein Dimerization in Cell-Derived Lipid Membranes Measured by FRET with MC Simulations
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F20%3A00523933" target="_blank" >RIV/61388963:_____/20:00523933 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11110/20:10422991 RIV/00216208:11310/20:10422991
Result on the web
<a href="https://www.sciencedirect.com/science/article/pii/S0006349520302605" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0006349520302605</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.bpj.2020.03.011" target="_blank" >10.1016/j.bpj.2020.03.011</a>
Alternative languages
Result language
angličtina
Original language name
Membrane Protein Dimerization in Cell-Derived Lipid Membranes Measured by FRET with MC Simulations
Original language description
Many membrane proteins are thought to function as dimers or higher oligomers, but measuring membrane protein oligomerization in lipid membranes is particularly challenging. Förster resonance energy transfer (FRET) and fluorescence cross-correlation spectroscopy are noninvasive, optical methods of choice that have been applied to the analysis of dimerization of single-spanning membrane proteins. However, the effects inherent to such two-dimensional systems, such as the excluded volume of polytopic transmembrane proteins, proximity FRET, and rotational diffusion of fluorophore dipoles, complicate interpretation of FRET data and have not been typically accounted for. Here, using FRET and fluorescence cross-correlation spectroscopy, we introduce a method to measure surface protein density and to estimate the apparent Förster radius, and we use Monte Carlo simulations of the FRET data to account for the proximity FRET effect occurring in confined two-dimensional environments. We then use FRET to analyze the dimerization of human rhomboid protease RHBDL2 in giant plasma membrane vesicles. We find no evidence for stable oligomers of RHBDL2 in giant plasma membrane vesicles of human cells even at concentrations that highly exceed endogenous expression levels. This indicates that the rhomboid transmembrane core is intrinsically monomeric. Our findings will find use in the application of FRET and fluorescence correlation spectroscopy for the analysis of oligomerization of transmembrane proteins in cell-derived lipid membranes.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10610 - Biophysics
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Biophysical Journal
ISSN
0006-3495
e-ISSN
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Volume of the periodical
118
Issue of the periodical within the volume
8
Country of publishing house
US - UNITED STATES
Number of pages
15
Pages from-to
1861-1875
UT code for WoS article
000528253800008
EID of the result in the Scopus database
2-s2.0-85082663310