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Titolo:
Pseudomonas aeruginosa protease IV enzyme assays and comparison to other pseudomonas proteases
Autore:
Caballero, AR; Moreau, JM; Engel, LS; Marquart, ME; Hill, JM; OCallaghan, RJ;
Indirizzi:
Louisiana State Univ, Hlth Sci Ctr, Dept Microbiol Immunol & Parasitol, New Orleans, LA 70112 USA Louisiana State Univ New Orleans LA USA 70112 , New Orleans, LA 70112 USA Louisiana State Univ, Ctr Eye, Dept Ophthalmol, New Orleans, LA 70112 USA Louisiana State Univ New Orleans LA USA 70112 , New Orleans, LA 70112 USA
Titolo Testata:
ANALYTICAL BIOCHEMISTRY
fascicolo: 2, volume: 290, anno: 2001,
pagine: 330 - 337
SICI:
0003-2697(20010315)290:2<330:PAPIEA>2.0.ZU;2-Y
Fonte:
ISI
Lingua:
ENG
Soggetto:
ALKALINE PROTEASE; BACTERIAL KERATITIS; POLYACRYLAMIDE GELS; CLINICAL STRAINS; VIRULENCE FACTOR; CYSTIC-FIBROSIS; LASA GENE; ELASTASE; PROTEINS; PURIFICATION;
Keywords:
alkaline protease; elastase A; elastase B; protease IV; Pseudomonas;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
52
Recensione:
Indirizzi per estratti:
Indirizzo: O'Callaghan, RJ Louisiana State Univ, Hlth Sci Ctr, Dept Microbiol Immunol& Parasitol, 1901 Perdido St, New Orleans, LA 70112 USA Louisiana State Univ 1901 Perdido St New Orleans LA USA 70112
Citazione:
A.R. Caballero et al., "Pseudomonas aeruginosa protease IV enzyme assays and comparison to other pseudomonas proteases", ANALYT BIOC, 290(2), 2001, pp. 330-337

Abstract

Pseudomonas aeruginosa secretes multiple proteases that have been implicated as virulence factors and the detection of each specific enzyme can be difficult to determine. Unlike the three Pseudomonas enzymes that have been well characterized (elastase A, elastase B, and alkaline protease), the activity of protease TV in multiple assays has yet to be described. This study defines new assays for Pseudomonas proteases and compares protease IV activity to the activities of elastase A, elastase B, and alkaline protease. Sixin vitro assays were studied: zymography, elastin congo red assay, staphylolytic assay, colorimetric peptide assay, solid-phase colorimetric peptide assay, and poly-L-lysine degradation. Casein zymography distinguished protease IV from elastase B and alkaline protease, and gelatin zymography differentiated all four proteases. The elastin congo red assay detected mainly elastase B while the staphylolytic assay was specific for elastase A. Protease IV activity was assayed specifically by the colorimetric assay and two new assays, the solid-phase colorimetric assay and degradation of poly-L-lysine in the presence of EDTA. Alkaline protease could be specifically assayedby poly-L-lysine degradation in the presence of N-alpha -p-to-syl-L-lysinechloromethyl ketone. The results identified three specific assays for protease TV, a new assay specific for alkaline protease, and showed that protease TV has a distinct enzymatic specificity relative to the three other Pseudomonas proteases. (C) 2001 Academic Press.

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Documento generato il 01/12/20 alle ore 19:04:51