Proteolytic processing of a secreted glycoprotein and O-glycosylation of mannoproteins are affected in the N-glycosylation mutant Saccharomyces cerevisiae ldb1.

摘要

In a previous work [P.I. Manas, I. Olivero, M. Avalos, L.M. Hernandez, Glycobiology, 7 (1997) 487-497], we described the isolation and characterization of the Saccharomyces cerevisiae ldb1 mutant which is affected in several steps of the N-glycosylation of mannoproteins probably due to a malfunction of the Golgi apparatus. Here, we found that two further functions assigned to the Golgi cisternae are also affected in the mutant: proteolytic processing of a secreted protein and O-glycosylation. We found that around 70% of the exoglucanase activity that is secreted into the culture medium by ldb1 bears an extra tetrapeptide in its NH2-terminus due to incomplete proteolytic processing. The O-linked oligosaccharides from ldb1 mnn1 were indistinguishable from those synthesized by the parental strain mnn1. However, when the O-oligosaccharides from the wild type and ldb1 were compared, we found a significant decrease in the tetrasaccharide in the latter, as well as a concomitant increase in the disaccharide, suggesting a defect in the Kre2p/Mnt1p involved in the transfer of the third mannose of these residues. Copyright 1998 Elsevier Science B.V.