title: Proteinases of Proteus spp.: purification, properties, and detection in urine of infected patients.
abstract: The proteinases secreted by pathogenic strains of Proteus mirabilis, P. vulgaris biotype 2, P. vulgaris biotype 3, and P. penneri were purified with almost 100% recovery by affinity chromatography on phenyl-Sepharose followed by anion-exchange chromatography. The proteinase purified from the urinary tract pathogen P. mirabilis, which we had previously shown to degrade immunoglobulins A and G, appeared as a composite of a single band and a double band (53 and 50 kDa, respectively) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The other Proteus proteinases had similar patterns but slightly different mobilities. In each case all proteinase activity in culture supernatants was demonstrated by gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be associated with only the triple-band complex; all three bands were proteolytically active. The P. mirabilis proteinase was resistant to inhibitors of both serine and thiol proteinases but strongly inhibited by metal chelators, although it was not affected by phosphoramidon, an inhibitor of the thermolysin group of bacterial metalloproteinases. Active proteinase was detected in urine samples from P. mirabilis-infected patients; this is consistent with our detection of immunoglobulin A fragments of a size suggestive of P. mirabilis proteinase activity.
authors.affiliation: Department of Pathology, Dundee University Medical School, Ninewells Hospital, Scotland.
authors.forename: L M
authors.initials: LM
authors.lastname: Loomes
authors.forename: B W
authors.initials: BW
authors.lastname: Senior
authors.forename: M A
authors.initials: MA
authors.lastname: Kerr
mesh_terms: D002846:Chromatography, Affinity; D010450:Endopeptidases / Q000302:isolation & purification* / Q000652:urine; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007070:Immunoglobulin A / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D008297:Male; D011512:Proteus Infections / Q000201:enzymology* / Q000652:urine; D011513:Proteus mirabilis / Q000201:enzymology*
chemical_list: D007070:Immunoglobulin A; D007074:Immunoglobulin G; D010450:Endopeptidases