Inhibition of cell adhesion in Dictyostelium discoideum by tunicamycin is prevented by leupeptin.
The carbohydrate requirement for cell adhesion of aggregation-competent cells of Dictyostelium discoideum has been examined by use of a selective glycosylation inhibitor of N-glycosyl protein, tunicamycin (TM). TM completely inhibited EDTA-stable cell adhesion and glycosylation of some membrane glycoproteins in aggregation-competent cells of D. discoideum (Yamada, H., et al. (1982) J. Biochem. 92, 399-406). The present study showed that the inhibition of EDTA-stable cell adhesion by TM was prevented significantly when the cells were treated with TM in the presence of a protease inhibitor, leupeptin (LP), whereas the inhibition of glycosylation by TM was not prevented. The cell extract of aggregation-competent cells contained acid proteases, and LP strongly inhibited acid protease from D. discoideum in vitro. On analysis by SDS-polyacrylamide gel electrophoresis (PAGE), many protein bands present in the membrane fraction of control cells disappeared or decreased on TM treatment of the cells in the absence of LP, however, some of these proteins were restored when the cells were treated with TM in the presence of LP. These results strongly support an idea that EDTA-stable cell adhesion characteristic to aggregation-competent cells is mediated by glycoproteins with asparagine-linked carbohydrate. However, the requirement for the carbohydrate moiety of the glycoprotein in cell adhesion appears to be indirect in that it acts to protect the protein moiety from proteolytic degradation.
D016282:Aspartic Acid Endopeptidases; D002448:Cell Adhesion / Q000187:drug effects; D004023:Dictyostelium / Q000166:cytology*; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology*; D009842:Oligopeptides / Q000494:pharmacology*; D014415:Tunicamycin / Q000494:pharmacology*
D007976:Leupeptins; D009842:Oligopeptides; D014415:Tunicamycin; D004492:Edetic Acid; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; D007930:Leucine; C032854:leupeptin; D005944:Glucosamine