Evaluation of P1' substrate specificity of staphylococcal SplB protease.

Katarzyna Pustelny; Natalia Stach; Benedykt Wladyka; Adam Dubin; Grzegorz Dubin

doi:10.18388/abp.2014_1937

Katarzyna Pustelny Department of Analytical Biochemistry and Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.;
Natalia Stach Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.;
Benedykt Wladyka Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Polandi and Malopolska Centre of Biotechnology, Kraków, Poland.;
Adam Dubin Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.;
Grzegorz Dubin Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Polan and Malopolska Centre of Biotechnology, Kraków, Poland.;

DOI: https://doi.org/10.18388/abp.2014_1937

Abstract

Staphylococcus aureus is a dangerous human pathogen characterized by growing antibiotic resistance. Virulence of S. aureus relies on a variety of secreted and cell surface associated virulence factors among which certain proteolytic enzymes play an important role. Amid staphylococcal extracellular proteases, those encoded by the spl operon remain poorly characterized, both in terms of enzymology and their physiological role. Initial data demonstrated that Spl proteases exhibit restricted substrate specificity. This study describes development of convenient protein FRET substrates for SplB protease and characterization of the substrate preference of the protease at the P1' position. Kinetic data on hydrolysis of a panel of substrates substituted at the said position is provided.