Glycosylation affects translocation of integrin, Src, and caveolin into or out of GEM.
Endogenous GM3 synthesis and full N-glycosylation in membrane receptors occurred in "4-epimerase-less" ldlD (Krieger's CHO mutant) cells cultured in Gal-containing medium, whereby components of detergent-insoluble, low-density, buoyant membrane fraction, termed "glycolipid-enriched microdomain (GEM)," varied significantly by translocation into or out of GEM. Integrins alpha3 and alpha5 were translocated into GEM in the presence of 0.5 or 0.25% Triton X-100, particularly in the absence of Gal, whereby integrins are underglycosylated and GlcCer is the major glycolipid component in GEM. Src family kinase was translocated into and enriched in GEM fractions when prepared in 0.5 or 0.25% Triton X-100 from cells grown in Gal-containing medium, whereby GM3 synthesis is induced. In contrast, caveolin is highly enriched in GEM when GM3 synthesis does not occur, and is translocated into high-density membrane fraction when GM3 synthesis occurs. The results suggest that levels of key molecules controlling cell adhesion and signaling are defined by translocation into or out of GEM, which depends on glycosylation state.
Kazui
A
A
Pacific Northwest Research Institute, University of Washington, 720 Broadway, Seattle, Washington 98122-4327, USA.
Ono
M
M
Handa
K
K
Hakomori
S
S
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D016466:CHO Cells; D051242:Caveolin 1; D022461:Caveolins; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism*; D006224:Cricetinae; D005679:G(M3) Ganglioside / Q000378:metabolism; D005690:Galactose / Q000378:metabolism / Q000494:pharmacology; D017353:Gene Deletion; D005963:Glucosylceramides / Q000378:metabolism; D006017:Glycolipids / Q000032:analysis*; D006031:Glycosylation / Q000187:drug effects; D039422:Integrin alpha3; D039482:Integrin alpha5; D016023:Integrins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D017830:Octoxynol / Q000378:metabolism / Q000494:pharmacology; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism; D012995:Solubility; D014534:UDPglucose 4-Epimerase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism*
D015703:Antigens, CD; D051242:Caveolin 1; D022461:Caveolins; D005679:G(M3) Ganglioside; D005963:Glucosylceramides; D006017:Glycolipids; D039422:Integrin alpha3; D039482:Integrin alpha5; D016023:Integrins; D008565:Membrane Proteins; D017830:Octoxynol; D016392:Proto-Oncogene Proteins pp60(c-src); D019061:src-Family Kinases; D014534:UDPglucose 4-Epimerase; D005690:Galactose