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PMID: 20051050 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Gangliosides and beta1-integrin are required for caveolae and membrane domains.

Traffic (Copenhagen, Denmark) ·Vol. 11 ·No. 3 ·2010-03-00 ·Pages 348-60

Singh RD, Marks DL, Holicky EL, Wheatley CL, Kaptzan T, Sato SB, Kobayashi T, Ling K, Pagano RE

Abstract

Caveolae are plasma membrane domains involved in the uptake of certain pathogens and toxins. Internalization of some cell surface integrins occurs via caveolae suggesting caveolae may play a crucial role in modulating integrin-mediated adhesion and cell migration. Here we demonstrate a critical role for gangliosides (sialo-glycosphingolipids) in regulating caveolar endocytosis in human skin fibroblasts. Pretreatment of cells with endoglycoceramidase (cleaves glycosphingolipids) or sialidase (modifies cell surface gangliosides and glycoproteins) selectively inhibited caveolar endocytosis by >70%, inhibited the formation of plasma membrane domains enriched in sphingolipids and cholesterol ('lipid rafts'), reduced caveolae and caveolin-1 at the plasma membrane by approximately 80%, and blunted activation of beta1-integrin, a protein required for caveolar endocytosis in these cells. These effects could be reversed by a brief incubation with gangliosides (but not with asialo-gangliosides or other sphingolipids) at 10 degrees C, suggesting that sialo-lipids are critical in supporting caveolar endocytosis. Endoglycoceramidase treatment also caused a redistribution of focal adhesion kinase, paxillin, talin, and PIP Kinase Igamma away from focal adhesions. The effects of sialidase or endoglycoceramidase on membrane domains and the distribution of caveolin-1 could be recapitulated by beta1-integrin knockdown. These results suggest that both gangliosides and beta1-integrin are required for maintenance of caveolae and plasma membrane domains.

MeSH Terms
Caveolae/metabolism Caveolin 1/metabolism Endocytosis Fibroblasts/metabolism Focal Adhesion Protein-Tyrosine Kinases/metabolism Gangliosides/metabolism Glycoside Hydrolases/pharmacology Humans Integrin beta1/metabolism Membrane Microdomains/metabolism Neuraminidase/pharmacology Paxillin/metabolism Skin/metabolism Talin/metabolism
Chemicals
Caveolin 1 Gangliosides Integrin beta1 Paxillin Talin Focal Adhesion Protein-Tyrosine Kinases Glycoside Hydrolases endoglycoceramidase Neuraminidase
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Singh Raman Deep
Departments of Medicine, Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
Marks David L
Holicky Eileen L
Wheatley Christine L
Kaptzan Tatiana
Sato Satoshi B
Kobayashi Toshihide
Ling Kun
Pagano Richard E
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Article Info
Journal
Traffic (Copenhagen, Denmark)
Abbr.
Traffic
ISSN
1600-0854
DOI
10.1111/j.1600-0854.2009.01022.x
Published
2010-03-00
Epub
2009-00-03
Pages
348-60
Language
English
Region
England
NLM ID
100939340
PMCID
PMC2852475
Subset
IM
Grants
NIGMS NIH HHS · R01 GM022942 · United States
NIGMS NIH HHS · GM-22942 · United States
NIGMS NIH HHS · R01 GM060934-10 · United States
NIGMS NIH HHS · R01 GM060934 · United States
NIGMS NIH HHS · GM-60934 · United States
NIGMS NIH HHS · R01 GM022942-35 · United States
External Links
PubMed source DOI: 10.1111/j.1600-0854.2009.01022.x PMC Full Text View journal details
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