title: N-deglycosylation of human complement component C9 reduces its hemolytic activity.
abstract: The effect of enzymatic deglycosylation of human complement component C9 on its hemolytic activity was investigated. Treatment of native C9 (Mr 71,000) with glyocpeptidase F (PNGase F) results in a stepwise decrease of the mol. wt. The formation of an Mr 67,000 peptide which is further converted to Mr 63,000 suggests that there are two N-linked carbohydrate chains per C9 polypeptide. Removal of approximately 88% of the N-linked oligosaccharides results in 80% reduction of the hemolytic activity (CH50). The completely N-deglycosylated Mr 63,000 peptide contains a remaining amount of 25% of the total carbohydrates of native C9. These glycans are assumed to be O-linked and predominantly attached to the C9a part of C9. The electrophoretic mobility of C9 is not affected by endoglycosidase F or H treatments revealing that the two N-linked glycans are of the tri- or tetra-antennary complex type. Cleavage of terminal sialic acids from native C9 by neuraminidase results in an Mr 67,000 product with nearly unaltered hemolytic activity. In contrast to other glycoproteins in which deglycosylation remained without major effects on their functional activity, our findings suggest that the N-linked carbohydrates are required for full expression of hemolytic activity of C9.
authors.affiliation: Institute of Immunology, University of Heidelberg, F.R.G.
authors.forename: R
authors.initials: R
authors.lastname: Kontermann
authors.forename: E W
authors.initials: EW
authors.lastname: Rauterberg
mesh_terms: D000581:Amidohydrolases / Q000494:pharmacology; D003186:Complement C9 / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis / Q000276:immunology*; D006801:Humans; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D013329:Structure-Activity Relationship
chemical_list: D003186:Complement C9; D009844:Oligosaccharides; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase