Dynamic Organization of Myristoylated Src in the Live Cell Plasma Membrane

The Journal of Physical Chemistry. B
Adam W SmithJay T Groves

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 i...Continue Reading

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Citations

Aug 31, 2016·Journal of the American Chemical Society·Philip W FowlerPeter D Olmsted
Apr 16, 2020·International Journal of Molecular Sciences·Giulia BagnatoBarbara Peruzzi
Jun 18, 2020·International Journal of Molecular Sciences·Chakia J McClendon, W Todd Miller

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