Effects of N-3554S, a polyprenyl phosphate, on B16-F10 mouse melanoma cells.
N-3554S, an optically active S-isomer of alpha-dihydrodecaprenyl phosphate, reduced the tumorigenicity of cultured B16-F10 mouse melanoma cells probably by affecting protein N-glycosylation. Accordingly, membrane glycoprotein samples were prepared from the melanoma cells cultured with or without N-3554S, and amounts and structures of N-linked sugar chains were determined. Analyses of the N-linked oligosaccharides released by hydrazinolysis from these samples and reduced with NaB3H4 revealed that the N-3554S-treated cells contain 1.5-1.8 times as much oligosaccharides as the control cells, and the relative amounts of high-mannose-type and bi-, tri- and tetra-antennary complex-type sugar chains are almost the same between two samples. Western blot analysis, however, showed that binding of L-PHA, which binds to oligosaccharides with the GlcNAc beta 1-->6(GlcNAc beta 1-->2)Man structure, is significantly reduced in 90 K, 96 K, 140 K, 155 K and 180 K glycoproteins in N-3554S-treated cells. Immunoblot analysis showed that the 140 K glycoprotein could be a fibronectin receptor. It was also shown that N-3554S treatment enhances the adhesiveness of the cells to fibronectin. These results indicate that N-3554S affects N-glycosylation of membrane glycoproteins and alters the cell surface properties of B16-F10 cells.
Shirane
K
K
Pharmaceutical Research Center, Nisshin Flour Milling Co., Ltd., Saitama, Japan.
Furukawa
K
K
Fukuchi
K
K
Yamazaki
H
H
Tsuji
M
M
Okamoto
Y
Y
D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D005353:Fibronectins / Q000378:metabolism; D006026:Glycoside Hydrolases; D006031:Glycosylation / Q000187:drug effects; D008545:Melanoma / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis; D011106:Polyisoprenyl Phosphates / Q000494:pharmacology*; D013237:Stereoisomerism; D014407:Tumor Cells, Cultured / Q000378:metabolism
D005353:Fibronectins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011106:Polyisoprenyl Phosphates; C048155:dihydrodecaprenyl phosphate; D006026:Glycoside Hydrolases