Home LiteratureArticle Details
PMID: 2110137 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Purification and characterization of an active fragment of the LasA protein from Pseudomonas aeruginosa: enhancement of elastase activity.

Journal of bacteriology ·Vol. 172 ·No. 5 ·1990-05-00 ·Pages 2236-40

Peters JE, Galloway DR

Abstract

A 22-kilodalton protein purified from the culture supernatant fraction of Pseudomonas aeruginosa (strains PA220 and PAO1) was found to enhance the elastolytic activity of purified P. aeruginosa elastase. N-terminal sequence analysis identified the protein as a fragment of the lasA gene product (P.A. Schad and B.H. Iglewski, J. Bacteriol. 170:2784-2789, 1988). However, comparative analysis with the reported LasA sequence indicated that the purified LasA fragment is longer than the deduced sequence reported. The purified LasA fragment had minimal elastolytic and proteolytic activity and did not enhance the proteolytic activity of purified elastase, yet enhanced the elastolytic activity more than 25-fold. The LasA fragment was found to also enhance the elastolytic activities of thermolysin, human neutrophil elastase, and proteinase K. The results presented here suggest that the LasA protein interacts with the elastin substrate rather than modifying elastase.

MeSH Terms
Bacterial Proteins/isolation & purification,metabolism Chromatography, Ion Exchange Electrophoresis, Polyacrylamide Gel Kinetics Metalloendopeptidases Molecular Weight Pancreatic Elastase/isolation & purification,metabolism Peptide Hydrolases/metabolism Pseudomonas aeruginosa/enzymology
Chemicals
Bacterial Proteins Peptide Hydrolases Pancreatic Elastase Metalloendopeptidases staphylolytic protease
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Peters J E
Department of Microbiology, Ohio State University, Columbus 43210-1292.
Galloway D R
References (26)
26 references, click to expand
  1. The role of the elastase of Pseudomonas aeruginosa in experimental infection.
    Exp Mol Pathol. 1965 Dec;4(6):567-75 PMID: 4954700
  2. Structural gene and complete amino acid sequence of Pseudomonas aeruginosa IFO 3455 elastase.
    J Bacteriol. 1989 Mar;171(3):1698-704 PMID: 2493453
  3. [Elastase in pseudomonas and aeromonas].
    Zentralbl Bakteriol Orig A. 1972 May;220(1):435-42 PMID: 4145620
  4. Elastase of Pseudomonas aeruginosa: inactivation of complement components and complement-derived chemotactic and phagocytic factors.
    Infect Immun. 1974 Jul;10(1):128-35 PMID: 4210424
  5. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  6. Pseudomonas protease. Purification, partial characterization, and its effect on collagen, proteoglycan, and rabbit corneas.
    Invest Ophthalmol Vis Sci. 1977 Jun;16(6):488-97 PMID: 405343
  7. Passive protection by antitoxin in experimental Pseudomonas aeruginosa burn infections.
    Infect Immun. 1977 Dec;18(3):596-602 PMID: 412786
  8. Protease and elastase of Pseudomonas aeruginosa: inactivation of human plasma alpha 1-proteinase inhibitor.
    Infect Immun. 1979 Apr;24(1):188-93 PMID: 110691
  9. Chromosomal genetics of Pseudomonas.
    Microbiol Rev. 1979 Mar;43(1):73-102 PMID: 111024
  10. Isolation and characterization of Pseudomonas aeruginosa PAO mutant that produces altered elastase.
    J Bacteriol. 1980 Jun;142(3):836-42 PMID: 6769912
  11. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  12. [Extracellular toxins of Pseudomonas aeruginosa. II. Effect of two proteases on human immunoglobulins IgG, IgA and secretory IgA (author's transl)].
    Zentralbl Bakteriol A. 1981 Mar;249(1):89-98 PMID: 6791405
  13. Protease phenotypes of Pseudomonas aeruginosa isolated from patients with cystic fibrosis.
    J Clin Microbiol. 1983 Jan;17(1):55-9 PMID: 6402520
  14. Experimental studies of the pathogenesis of infections owing to Pseudomonas aeruginosa: elastase, an IgG protease.
    Can J Microbiol. 1984 Sep;30(9):1118-24 PMID: 6439405
  15. Analytical isoelectric focusing using a high-voltage vertical slab polyacrylamide gel system.
    Anal Biochem. 1984 Nov 1;142(2):421-36 PMID: 6528977
  16. Cloning and characterization of elastase genes from Pseudomonas aeruginosa.
    J Bacteriol. 1987 Jun;169(6):2691-6 PMID: 3034864
  17. Elastolytic activity of Pseudomonas aeruginosa elastase.
    Biochim Biophys Acta. 1989 May 1;995(3):285-90 PMID: 2495818
  18. PRODUCTION OF ELASTASE AND PROTEINASE BY PSEUDOMONAS AERUGINOSA.
    J Bacteriol. 1964 Sep;88:745-57 PMID: 14208514
  19. PSEUDOMONAS AERUGINOSA ELASTASE. ISOLATION, CRYSTALLIZATION, AND PRELIMINARY CHARACTERIZATION.
    J Biol Chem. 1965 Aug;240:3295-304 PMID: 14321366
  20. Biochemical and pathologic evidence for proteolytic destruction of lung connective tissue in cystic fibrosis.
    Am Rev Respir Dis. 1985 Sep;132(3):529-35 PMID: 3898942
  21. Cloning and transcriptional regulation of the elastase lasA gene in mucoid and nonmucoid Pseudomonas aeruginosa.
    J Bacteriol. 1987 Mar;169(3):1349-51 PMID: 3102460
  22. Primary structures of bovine elastin a, b, and c deduced from the sequences of cDNA clones.
    J Biol Chem. 1987 Apr 25;262(12):5755-62 PMID: 3032943
  23. Activation of an elastase precursor by the lasA gene product of Pseudomonas aeruginosa.
    J Bacteriol. 1987 Oct;169(10):4532-9 PMID: 2820931
  24. Nucleotide sequence and expression in Escherichia coli of the Pseudomonas aeruginosa lasA gene.
    J Bacteriol. 1988 Jun;170(6):2784-9 PMID: 2836371
  25. Molecular characterization and nucleotide sequence of the Pseudomonas aeruginosa elastase structural gene.
    J Bacteriol. 1988 Sep;170(9):4309-14 PMID: 2842313
  26. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
1990-05-00
Pages
2236-40
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC208853
Subset
IM
Grants
NIAID NIH HHS · AI00876-01 · United States
NIAID NIH HHS · AI123429-02 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com