Cargando…
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...
Autores principales: | , , , , , |
---|---|
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 |
_version_ | 1782409789244964864 |
---|---|
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. |
format | Online Article Text |
id | pubmed-4710234 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT sahasuvrajit diffusionofgpianchoredproteinsisinfluencedbytheactivityofdynamiccorticalactin AT leeilhyung diffusionofgpianchoredproteinsisinfluencedbytheactivityofdynamiccorticalactin AT polleyanirban diffusionofgpianchoredproteinsisinfluencedbytheactivityofdynamiccorticalactin AT grovesjayt diffusionofgpianchoredproteinsisinfluencedbytheactivityofdynamiccorticalactin AT raomadan diffusionofgpianchoredproteinsisinfluencedbytheactivityofdynamiccorticalactin AT mayorsatyajit diffusionofgpianchoredproteinsisinfluencedbytheactivityofdynamiccorticalactin |