Decreased adenylate cyclase activation by helodermin and PGE1 in the lectin-resistant variant Wa4 of the mouse melanoma cell line B16.
We examined the cultured mouse melanoma cell line B16 (clone F1) and its wheat germ agglutinin-resistant variant Wa4 that suffers from abnormal protein glycosylation (a high fucose:sialic acid ratio in glycoproteins). In both cell lines the adenylate cyclase system was endowed with a functional guanine nucleotide binding protein Gs and was efficiently coupled to alpha-MSH receptors. In the B16 cell line F1 studied we also observed an efficient stimulation of adenylate cyclase activity by helodermin, VIP and the VIP analogue [acetyl-His1]VIP, and also by PGE1. In membranes from the lectin-resistant variant Wa4, the stimulations by VIP-like peptides and by PGE1 were reduced by 60% and 50%, respectively, while the stimulation by alpha-MSH remained normal. As other components of the adenylate cyclase system (Gs site, catalytical unit) appeared unchanged in the Wa4 variant, we conclude that impaired glycosylation essentially affected the number of both VIP-like peptide receptors and PGE1 receptors.
Damien
C
C
Department of Biochemistry and Nutrition, Medical School, Université Libre de Bruxelles, Brussels, Belgium.
Robberecht
P
P
Hooghe
R
R
De Neef
P
P
Christophe
J
J
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000527:Alprostadil / Q000494:pharmacology*; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002772:Cholera Toxin / Q000494:pharmacology; D002999:Clone Cells; D004789:Enzyme Activation / Q000187:drug effects; D036341:Intercellular Signaling Peptides and Proteins; D008546:Melanoma, Experimental / Q000201:enzymology*; D051379:Mice; D010455:Peptides / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology; D014909:Wheat Germ Agglutinins / Q000494:pharmacology*; D000521:alpha-MSH / Q000494:pharmacology
D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D014909:Wheat Germ Agglutinins; D014660:Vasoactive Intestinal Peptide; D000521:alpha-MSH; C040442:heliodermin; D002772:Cholera Toxin; D000262:Adenylyl Cyclases; D000527:Alprostadil