Analysis of individual azurocidin N-glycosylation sites in regard to its secretion by insect cells, susceptibility to proteolysis and antibacterial activity.
Azurocidin is an inactive serine protease homolog with primary sequence similarity to neutrophil elastase, cathepsin G, and proteinase 3. The aim of this study was to investigate possible consequences of differential glycosylation of azurocidin in regard to its secretion, protein stability as measured by susceptibility to proteolysis, and antibacterial activity. Site-directed mutagenesis was employed to generate mutant azurocidin variants lacking individual N-glycosylation sites. Our results show that N-linked glycans may play a role in proper azurocidin folding and subsequent secretion by insect cells. We also demonstrate that N-linked glycosylation contributes to azurocidin stability by protecting it from proteolysis. The lack of N-glycosylation at individual sites does not significantly influence the azurocidin antibacterial activity.
Indyk
Katarzyna
K
Laboratory of Biochemistry, Faculty of Biotechnology, University of Wrocław, Wrocław, Poland.
Olczak
Teresa
T
Ciuraszkiewicz
Justyna
J
Watorek
Wiesław
W
Olczak
Mariusz
M
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D002460:Cell Line; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development; D006031:Glycosylation; D006868:Hydrolysis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism
C062991:AZU1 protein, human; D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; D014357:Trypsin