title: A mouse B16 melanoma mutant deficient in glycolipids.
abstract: Mouse B16 melanoma cell line, GM-95 (formerly designated as MEC-4), deficient in sialyllactosylceramide was examined for its primary defect. Glycolipids from the mutant cells were analyzed by high-performance TLC. No glycolipid was detected in GM-95 cells, even when total lipid from 10(7) cells was analyzed. In contrast, the content of ceramide, a precursor lipid molecule of glycolipids, was normal. Thus, the deficiency of glycolipids was attributed to the first glucosylation step of ceramide. The ceramide glucosyltransferase (EC 2.4.1.80) activity was not detected in GM-95 cells. There was no significant difference of sialyllactosylceramide synthase activity, however, between GM-95 and the parental cells. The deficiency of glycolipids in GM-95 cells was associated with changes of the cellular morphology and growth rate. The parental cells showed irregular shapes and tended to overlap each other. On the other hand, GM-95 cells exhibited an elongated fibroblastic morphology and parallel arrangement. The population-doubling times of GM-95 and the parental cells in serum-free medium were 28 hr and 19 hr, respectively.
authors.affiliation: Laboratory for Glyco Cell Biology, Institute of Chemical and Physical Research (RIKEN), Saitama, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Ichikawa
authors.forename: N
authors.initials: N
authors.lastname: Nakajo
authors.forename: H
authors.initials: H
authors.lastname: Sakiyama
authors.forename: Y
authors.initials: Y
authors.lastname: Hirabayashi
mesh_terms: D000818:Animals; D002455:Cell Division; D016895:Culture Media, Serum-Free; D005455:Fluorescent Antibody Technique; D005679:G(M3) Ganglioside / Q000096:biosynthesis / Q000172:deficiency*; D005964:Glucosyltransferases / Q000172:deficiency* / Q000235:genetics; D037102:Lectins / Q000378:metabolism; D008055:Lipids / Q000032:analysis; D008546:Melanoma, Experimental / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D009154:Mutation
chemical_list: D016895:Culture Media, Serum-Free; D005679:G(M3) Ganglioside; D037102:Lectins; D008055:Lipids; D005964:Glucosyltransferases; C023246:ceramide glucosyltransferase