A neutral glycoprotease of Pasteurella haemolytica A1 specifically cleaves O-sialoglycoproteins

• Published in: Infection and Immunity Vol. 60; no. 1; pp. 56 - 62
• Main Authors: Abdullah, K.M. (Guelph-Waterloo Centre for Graduate Work in Chemistry, Ontario, Canada), Udoh, E.A, Shewen, P.E, Mellors, A
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-01-1992
• Subjects: ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; Ascorbic Acid - pharmacology; Bacteriology; Biological and medical sciences; Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Citrates - pharmacology; Citric Acid; Edetic Acid - pharmacology; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; GLICOPROTEINAS; Glycopeptides - pharmacology; Glycophorin - metabolism; GLYCOPROTEINE; Humans; Hydrolysis - drug effects; In Vitro Techniques; INHIBIDORES DE ENZIMAS; INHIBITEUR D'ENZYME; Mannheimia haemolytica - enzymology; Metabolism. Enzymes; Metalloendopeptidases - isolation & purification; Metalloendopeptidases - physiology; Microbiology; Neuraminidase - pharmacology; PASTEURELLA; PROPIEDADES FISICO-QUIMICAS; PROPRIETE PHYSICOCHIMIQUE; PROTEASAS; PROTEASE; PURIFICACION; PURIFICATION; Sialoglycoproteins - metabolism; Substrate Specificity; Sialic acid; Enzyme; MNS-System; Red blood cell; Pasteurellaceae; Sialoglycoproteins; Specificity; Enzymatic activity; O-Glucosidic bond; Plasma membrane; Bacteria; Metalloproteinase; Inhibition; Pasteurella haemolytica
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/iai.60.1.56-62.1992

A neutral metalloprotease with marked specificity for an O-sialoglycoprotein has been isolated from culture supernatants of Pasteurella haemolytica A1. The 35-kDa enzyme cleaves human erythrocyte glycophorin A, which is O glycosylated, but does not cleave N-glycosylated proteins or nonglycosylated proteins. Glycophorin A was cleaved when it was present in situ in erythrocyte ghost plasma membranes or when it was free in solution. The glycoprotease did not hydrolyze glycophorin A from which sialate residues had been removed by neuraminidase treatment. An immobilized preparation of the enzyme cleaved glycophorin A at several positions, with a major site of cleavage at Arg-31-Asp-32. The glycoprotease is inhibited by EDTA, citrate, and ascorbate, but inhibition appears to be due to the masking of metal ion activators rather than to their removal. The enzyme is not inhibited by phosphoramidon, an inhibitor of other bacterial neutral metalloproteases