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Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin

Molecular diffusion at the surface of living cells is believed to be predominantly driven by thermal kicks. However, there is growing evidence that certain cell surface molecules are driven by the fluctuating dynamics of cortical cytoskeleton. Using fluorescence correlation spectroscopy, we measure...

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Autores principales: Saha, Suvrajit, Lee, Il-Hyung, Polley, Anirban, Groves, Jay T., Rao, Madan, Mayor, Satyajit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710234/
https://www.ncbi.nlm.nih.gov/pubmed/26378258
http://dx.doi.org/10.1091/mbc.E15-06-0397
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author Saha, Suvrajit
Lee, Il-Hyung
Polley, Anirban
Groves, Jay T.
Rao, Madan
Mayor, Satyajit
author_facet Saha, Suvrajit
Lee, Il-Hyung
Polley, Anirban
Groves, Jay T.
Rao, Madan
Mayor, Satyajit
author_sort Saha, Suvrajit
collection PubMed
description Molecular diffusion at the surface of living cells is believed to be predominantly driven by thermal kicks. However, there is growing evidence that certain cell surface molecules are driven by the fluctuating dynamics of cortical cytoskeleton. Using fluorescence correlation spectroscopy, we measure the diffusion coefficient of a variety of cell surface molecules over a temperature range of 24–37°C. Exogenously incorporated fluorescent lipids with short acyl chains exhibit the expected increase of diffusion coefficient over this temperature range. In contrast, we find that GPI-anchored proteins exhibit temperature-independent diffusion over this range and revert to temperature-dependent diffusion on cell membrane blebs, in cells depleted of cholesterol, and upon acute perturbation of actin dynamics and myosin activity. A model transmembrane protein with a cytosolic actin-binding domain also exhibits the temperature-independent behavior, directly implicating the role of cortical actin. We show that diffusion of GPI-anchored proteins also becomes temperature dependent when the filamentous dynamic actin nucleator formin is inhibited. However, changes in cortical actin mesh size or perturbation of branched actin nucleator Arp2/3 do not affect this behavior. Thus cell surface diffusion of GPI-anchored proteins and transmembrane proteins that associate with actin is driven by active fluctuations of dynamic cortical actin filaments in addition to thermal fluctuations, consistent with expectations from an “active actin-membrane composite” cell surface.
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spelling pubmed-47102342016-01-20 Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin Saha, Suvrajit Lee, Il-Hyung Polley, Anirban Groves, Jay T. Rao, Madan Mayor, Satyajit Mol Biol Cell Articles Molecular diffusion at the surface of living cells is believed to be predominantly driven by thermal kicks. However, there is growing evidence that certain cell surface molecules are driven by the fluctuating dynamics of cortical cytoskeleton. Using fluorescence correlation spectroscopy, we measure the diffusion coefficient of a variety of cell surface molecules over a temperature range of 24–37°C. Exogenously incorporated fluorescent lipids with short acyl chains exhibit the expected increase of diffusion coefficient over this temperature range. In contrast, we find that GPI-anchored proteins exhibit temperature-independent diffusion over this range and revert to temperature-dependent diffusion on cell membrane blebs, in cells depleted of cholesterol, and upon acute perturbation of actin dynamics and myosin activity. A model transmembrane protein with a cytosolic actin-binding domain also exhibits the temperature-independent behavior, directly implicating the role of cortical actin. We show that diffusion of GPI-anchored proteins also becomes temperature dependent when the filamentous dynamic actin nucleator formin is inhibited. However, changes in cortical actin mesh size or perturbation of branched actin nucleator Arp2/3 do not affect this behavior. Thus cell surface diffusion of GPI-anchored proteins and transmembrane proteins that associate with actin is driven by active fluctuations of dynamic cortical actin filaments in addition to thermal fluctuations, consistent with expectations from an “active actin-membrane composite” cell surface. The American Society for Cell Biology 2015-11-05 /pmc/articles/PMC4710234/ /pubmed/26378258 http://dx.doi.org/10.1091/mbc.E15-06-0397 Text en © 2015 Saha et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Saha, Suvrajit
Lee, Il-Hyung
Polley, Anirban
Groves, Jay T.
Rao, Madan
Mayor, Satyajit
Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin
title Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin
title_full Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin
title_fullStr Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin
title_full_unstemmed Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin
title_short Diffusion of GPI-anchored proteins is influenced by the activity of dynamic cortical actin
title_sort diffusion of gpi-anchored proteins is influenced by the activity of dynamic cortical actin
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710234/
https://www.ncbi.nlm.nih.gov/pubmed/26378258
http://dx.doi.org/10.1091/mbc.E15-06-0397
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