Glycosylation Effect on Membrane Domain (GEM) Involved in Cell Adhesion and Motility: A Preliminary Note on Functional α3, α5-CD82 Glycosylation Complex in ldlD 14 Cells

doi:10.1006/bbrc.2000.4030

Biochemical and Biophysical Research Communications

Volume 279, Issue 3, 29 December 2000, Pages 744-750

https://doi.org/10.1006/bbrc.2000.4030 Get rights and content

Abstract

Laminin (LN)- or fibronectin (FN)-dependent adhesion in Krieger's ldlD 14 (D14) cells is enhanced significantly in the presence vs absence, of galactose (Gal), whereas LN- or FN-induced haptotactic cell motility is barely affected unless cells express CD82 by its gene transfection (cells termed D14/CD82). The effect of CD82 on LN- or FN-induced motility is based on its ability to associate with α3 or α5 integrin to form a complex associated with a low-density lipid membrane domain (termed GEM or GSD). Complex formation is greatly affected by N-glycosylation of both integrin and CD82, as well as by concurrent GM3 ganglioside synthesis. The effect of glycosylation on α5-CD82 complex was also studied in D14 cells expressing mutant CD82, defective in all three N-glycosylation sites. LN-induced motility was greatly inhibited, whereas FN-induced motility was enhanced, with complete N-glycosylation in D14/CD82 cells in Gal-added medium, whereby α5-CD82 complex formation did not occur or occurred at a minimal level. Both LN- and FN-induced motility were inhibited when N-glycosylation was impaired, or N-glycosylation of CD82 was deleted, whereby α5-CD82 complex formation occurred strongly. Thus, glycosylation profoundly affects interaction of integrin with CD82, leading to significant inhibition or promotion of cell motility.

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Citation Excerpt :
Moreover, the elevated level of α2-6 linked SA on the α5 integrin subunit in highly metastatic mouse hepatocarcinoma cells increased integrin α5β1-dependent cell adhesion to FN and facilitated FN-mediated phosphorylation of FAK [48]. On the other hand, α2–3,6 sialylation on the integrin α5 subunit caused the increase in FN-induced cell motility by abolished integrin α5β1 interaction with the metastasis suppressor CD82 [61]. In turn, the presence of α2-8 linked SA on the integrin α5 subunit of human G361 melanoma cells was crucial for integrin α5β1-dependent cell adhesion to FN and it was believed that these oligosialic acids stabilize integrin α5β1 conformation in a form exhibiting high affinity to the ligand [62].
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^☆: This work was supported by National Cancer Institute Grant CA80054 (to S.H.). M.O. was a recipient during 1998–2000 of a fellowship from the Department of Surgery, University of Tokyo School of Medicine, Japan.

^☆☆: Abbreviations used: co-IP, coimmunoprecipitation; co-IP'd, coimmunoprecipitated; ECM, extracellular matrix; FN, fibronectin; Gal, galactose; GSD, glycosignaling domain; GEM, glycosphingolipid-enriched microdomain; GSL, glycosphingolipid; LN, laminin; SM, sphingomyelin.

²: Present address: Department of Surgery, Jichi Medical School, Minamikawachi-machi, Tochigiken 329-0498, Japan.

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Regular ArticleGlycosylation Effect on Membrane Domain (GEM) Involved in Cell Adhesion and Motility: A Preliminary Note on Functional α3, α5-CD82 Glycosylation Complex in ldlD 14 Cells☆,☆☆

Abstract

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J. Biol. Chem.

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Curr. Opin. Cell Biol.

Cell

Meth. Cell Biol.

Biochem. Biophys. Res. Commun.

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Regular Article
Glycosylation Effect on Membrane Domain (GEM) Involved in Cell Adhesion and Motility: A Preliminary Note on Functional α3, α5-CD82 Glycosylation Complex in ldlD 14 Cells☆,☆☆