Fatty acid-dependent globotriaosyl ceramide receptor function in detergent resistant model membranes.
Glycosphingolipid (GSL) fatty acid strictly regulates verotoxin 1 (VT1) and the HIV adhesin, gp120 binding to globotriaosyl ceramide within Gb(3)/cholesterol detergent resistant membrane (DRM) vesicle constructs and in Gb(3) water-air interface monolayers in a similar manner. VT2 bound Gb(3)/cholesterol vesicles irrespective of fatty acid composition, but VT1 bound neither C18 nor C20Gb(3)vesicles. C18/C20Gb(3) were dominant negative in mixed Gb(3) fatty acid isoform vesicles, but including C24:1Gb(3) gave maximal binding. VT1 bound C18Gb(3) vesicles after cholesterol removal, but C20Gb(3)vesicles required sphingomyelin in addition for binding. HIV-1gp120 also bound C16, C22, and C24, but neither C18 nor C20Gb(3) vesicles. C18 and C20Gb(3) were, in mixtures without C24:1Gb(3), dominant negative for gp120 vesicle binding. Gp120/VT1bound C18 and C24:1Gb(3) mixtures, although neither isoform bound alone. Monolayer surface pressure measurement showed VT1, but not VT2, bound Gb(3) at cellular DRM surface pressures, and confirmed loss of VT1 and gp120 (but not VT2) specific C18Gb(3) binding. We conclude fatty-acid mediated fluidity within simple model GSL/cholesterol DRM can selectively regulate GSL carbohydrate-ligand binding.
Mahfoud
Radhia
R
Division of Molecular Structure and Function, Research Institute, The Hospital for Sick Children, Toronto, Ontario, Canada.
Manis
Adam
A
Lingwood
Clifford A
CA
D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D005227:Fatty Acids / Q000378:metabolism* / Q000494:pharmacology; D015699:HIV Envelope Protein gp120 / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008560:Membrane Fluidity / Q000187:drug effects; D011312:Pressure; D022641:Shiga Toxin 2 / Q000378:metabolism; D014281:Trihexosylceramides / Q000378:metabolism*
D003902:Detergents; D005227:Fatty Acids; D015699:HIV Envelope Protein gp120; D008024:Ligands; D022641:Shiga Toxin 2; D014281:Trihexosylceramides; C018549:globotriaosylceramide; D002784:Cholesterol