Effect of gangliosides on the distribution of a glycosylphosphatidylinositol-anchored protein in plasma membrane from CHO-K1 cells

CRESPO, PILAR MARIA; ZURITA, ADOLFO R.; DANIOTTI, JOSE LUIS

JOURNAL OF BIOLOGICAL CHEMISTRY

American Society for Biochemistry and Molecular Biology

Lugar: Bethesda, USA; Año: 2002 vol. 277 p. 44731 - 44731

0021-9258

lycosylphosphatidylinositol (GPI)-anchored proteinsare clustered mainly in sphingolipid-cholesterolmicrodomains of the plasma membrane. The distributionof GPI-anchored fusion yellow fluorescent protein(GPI-YFP) in the plasma membrane of Chinese hamsterovary (CHO)-K1 cells with different glycolipid compositionswas investigated. Cells depleted of glycosphingolipidsby inhibiting glucosylceramide synthase activityor cell lines expressing different gangliosides caused bystable transfection of appropriate ganglioside glycosyltransferasesor exposed to exogenous GM1 were transfectedwith GPI-YFP cDNA. The distribution of GPI-YFPfusion protein expressed at the plasma membrane wasstudied using the membrane-impermeable cross-linkingagent bis(sulfosuccinimidyl)suberate. Results indicatethat GPI-YFP forms clusters at the surface of cells expressingGM3, or cells depleted of glycolipids, or transfected