Dynamic organization of myristoylated Src in the live cell plasma membrane
Abstract
The spatial organization of lipid-anchored proteins in the plasma membrane directly influences cell signaling, but measuring such organization in situ is experimentally challenging. The canonical oncogene, c-Src, is a lipid anchored protein that plays a key role in integrin-mediated signal transduction within focal adhesions and cell–cell junctions. Because of its activity in specific plasma membrane regions, structural motifs within the protein have been hypothesized to play an important role in its subcellular localization. This study used a combination of time-resolved fluorescence fluctuation spectroscopy and super-resolution microscopy to quantify the dynamic organization of c-Src in live cell membranes. Pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE–FCCS) showed that a small fraction of c-Src transiently sorts into membrane clusters that are several times larger than the monomers. Photoactivated localization microscopy (PALM) confirmed that c-Src partitions into clusters with low probability and showed that the characteristic size of the clusters is 10–80 nm. Finally, time-resolved fluorescence anisotropy measurements were used to quantify the rotational mobility of c-Src to determine how it interacts with its local environment. Altogether, these results build a quantitative description of the mobility and clustering behavior of the c-Src nonreceptor tyrosine kinase in the live cell plasma membrane.
- Authors:
-
- Univ. of California, Berkeley, CA (United States); Univ. of Akron, Akron, OH (United States)
- Univ. of California, Berkeley, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1351532
- Report Number(s):
- LBNL-1004330
Journal ID: ISSN 1520-6106; ir:1004330
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry
- Additional Journal Information:
- Journal Volume: 120; Journal Issue: 5; Journal ID: ISSN 1520-6106
- Publisher:
- American Chemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 60 APPLIED LIFE SCIENCES
Citation Formats
Smith, Adam W., Huang, Hector H., Endres, Nicholas F., Rhodes, Christopher, and Groves, Jay T. Dynamic organization of myristoylated Src in the live cell plasma membrane. United States: N. p., 2016.
Web. doi:10.1021/acs.jpcb.5b08887.
Smith, Adam W., Huang, Hector H., Endres, Nicholas F., Rhodes, Christopher, & Groves, Jay T. Dynamic organization of myristoylated Src in the live cell plasma membrane. United States. https://doi.org/10.1021/acs.jpcb.5b08887
Smith, Adam W., Huang, Hector H., Endres, Nicholas F., Rhodes, Christopher, and Groves, Jay T. Fri .
"Dynamic organization of myristoylated Src in the live cell plasma membrane". United States. https://doi.org/10.1021/acs.jpcb.5b08887. https://www.osti.gov/servlets/purl/1351532.
@article{osti_1351532,
title = {Dynamic organization of myristoylated Src in the live cell plasma membrane},
author = {Smith, Adam W. and Huang, Hector H. and Endres, Nicholas F. and Rhodes, Christopher and Groves, Jay T.},
abstractNote = {The spatial organization of lipid-anchored proteins in the plasma membrane directly influences cell signaling, but measuring such organization in situ is experimentally challenging. The canonical oncogene, c-Src, is a lipid anchored protein that plays a key role in integrin-mediated signal transduction within focal adhesions and cell–cell junctions. Because of its activity in specific plasma membrane regions, structural motifs within the protein have been hypothesized to play an important role in its subcellular localization. This study used a combination of time-resolved fluorescence fluctuation spectroscopy and super-resolution microscopy to quantify the dynamic organization of c-Src in live cell membranes. Pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE–FCCS) showed that a small fraction of c-Src transiently sorts into membrane clusters that are several times larger than the monomers. Photoactivated localization microscopy (PALM) confirmed that c-Src partitions into clusters with low probability and showed that the characteristic size of the clusters is 10–80 nm. Finally, time-resolved fluorescence anisotropy measurements were used to quantify the rotational mobility of c-Src to determine how it interacts with its local environment. Altogether, these results build a quantitative description of the mobility and clustering behavior of the c-Src nonreceptor tyrosine kinase in the live cell plasma membrane.},
doi = {10.1021/acs.jpcb.5b08887},
journal = {Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry},
number = 5,
volume = 120,
place = {United States},
year = {Fri Jan 15 00:00:00 EST 2016},
month = {Fri Jan 15 00:00:00 EST 2016}
}
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Works referencing / citing this record:
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