Nucleotide sequence of the staphylocoagulase gene: its unique COOH-terminal 8 tandem repeats

Abstract

The entire staphylocoagulase gene of Staphylococcus aureus strain BB was cloned on a MboI restriction endonuclease fragment inserted into pAT153 plasmid vector. The staphylocoagulase was expressed in Escherichia coli, as judged by the formation of a fibrin halo on an agar plate containing rabbit plasma and bovine fibrinogen. We have determined the complete nucleotide sequence of the staphylocoagulase gene by the dideoxynucleotide chain termination method. The deduced amino acid sequence consisted of 715 residues including a signal peptide of 26 residues. Therefore, the predicted molecular weight of the mature protein was 77,337. This sequence was corroborated by reference to the amino acid compositions of 30 lysyl endopeptidase peptides and the sequences of 12 of these peptides isolated from the purified staphylocoagulase. The 5'-flanking region was found to contain a putative Shine-Dalgarno sequence and a putative "-10" element for transcription. The COOH-terminal stretch of 216 amino acids of staphylocoagulase was composed of 8 tandem repeats each consisting of 27 amino acid residues. The amino acid sequence of staphylocoagulase derived from strain BB showed 57% identity with that of the chymotryptic 43-kDa fragment of staphylocoagulase isolated previously from strain 213 (Kawabata, S., Miyata, To., Morita, T., Miyata, Ta., Iwanaga, S., & Igarashi, H. (1986) J. Biol. Chem. 261, 527-531).