Molecular cloning and biochemical characterisation of proteases from Staphylococcus epidermidis.
We report the complete coding sequence and the partial amino acid sequence (determined by chemical sequencing) of Staphylococcus epidermidis extracellular cysteine (Ecp) and serine (Esp) proteases. The first enzyme shows an extended sequence similarity to Staphylococcus aureus cysteine protease (staphopain) and the second one resembles the serine protease produced by that species. The region directly upstream of the sequence coding for the mature protein in both enzymes displays significant homology to the profragments encoded by sspB and sspA, respectively, thus suggesting that the characterised enzymes may also be produced as proproteins. Furthermore, we report some biological properties of the cysteine protease, contributing to a better understanding of its role as a possible virulence factor. The proteolytic activity of this enzyme was rapidly and efficiently inhibited by human alpha-2-macroglobulin; however, human kininogen as well as cystatins (A, C and D) were not inhibitory. Moreover, the protease was capable of inactivating, by limited proteolysis, both alpha-1-antitrypsin and HMW-kininogen, but neither alpha-1-antichymotrypsin nor antithrombin III.
Dubin
G
G
Institute of Molecular Biology, Jagiellonian University, Kraków, Poland.
Chmiel
D
D
Mak
P
P
Rakwalska
M
M
Rzychon
M
M
Dubin
A
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000378:metabolism; D010450:Endopeptidases / Q000737:chemistry*; D007704:Kininogens / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D012697:Serine Endopeptidases / Q000378:metabolism; D015843:Serpins / Q000096:biosynthesis / Q000235:genetics; D013212:Staphylococcus epidermidis / Q000201:enzymology* / Q000472:pathogenicity; D014774:Virulence
D001426:Bacterial Proteins; D007704:Kininogens; D015843:Serpins; D010450:Endopeptidases; D012697:Serine Endopeptidases; D003546:Cysteine Endopeptidases