A novel member of the thermolysin family, cloning and biochemical characterization of metalloprotease from Staphylococcus pseudintermedius

• Authors: Benedykt Wladyka , Michal Bista , Artur J Sabat , Emilia Bonar , Sabina Grzeszczuk , Waleria Hryniewicz , Adam Dubin
• Languages of publication: EN

Abstracts

EN

Thermolysins constitute a family of secreted bacterial metalloproteases expressed, among others, by several pathogens. Strains of Staphylococcus pseudintermedius isolated from diseased dogs and judged as protease-positive, by skim milk agar plate culture, were investigated for protease content. No proteolytic activity was detected when the bacteria were grown in regular liquid media. Unexpectedly, supplementation of the medium with calcium ions resulted in expression of a metalloprotease and profound changes in the profile of extracellular proteins. On the basis of homology to other staphylococcal metalloproteases, the nucleotide sequence of the gene encoding this protease (Pst) and its flanking regions was determined. The full-length pst codes for a protein with an open reading frame of 505 amino acids. The internal region contains the HEXXH catalytic domain that is conserved in members of the thermolysin family. Regardless of the presence of calcium in the medium, the expression of the protease gene was of the same intensity. This suggests that regulation of the metalloprotease production by calcium ions is at a post-transcriptional level. Isolates of S. pseudintermedius exhibit a proteolytic phenotype due to the metalloprotease expression, however only in presence of calcium ions, which most probably stabilize the structure of the protease.

Keywords

EN

staphylococcus; calcium; metalloprotease; pathogen

• Journal: Acta Biochimica Polonica
• Year: 2008
• Volume: 55
• Issue: 3
• Pages: 525-536

Dates

published

2008

received

2008-04-27

revised

2008-08-06

accepted

2008-09-01

(unknown)

2008-09-04

Contributors

author

Benedykt Wladyka

• Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Michal Bista

• Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Artur J Sabat

• Department of Epidemiology and Clinical Microbiology, National Institute of Medicines, Warszawa, Poland

author

Emilia Bonar

• Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Sabina Grzeszczuk

• Department of Epidemiology and Clinical Microbiology, National Institute of Medicines, Warszawa, Poland

author

Waleria Hryniewicz

• Department of Epidemiology and Clinical Microbiology, National Institute of Medicines, Warszawa, Poland

author

Adam Dubin

• Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

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