title: Novel self-cleavage activity of Staphylokinase fusion proteins: An interesting finding and its possible applications.
abstract: Staphylokinase (SAK) is reported to have a serine protease domain with no proteolytic activity unlike other plasminogen activators like tissue plasminogen activator (t-PA) and urokinase. A unique protease property of Staphylokinase was observed when SAK was expressed as a fusion protein in inducible Escherichia coli expression vectors. This finding was further investigated by cloning and expressing different SAK fusions, both native and N-terminal deletions, with fusion tags like glutathione S-transferase (GST) and signal sequence of SAK in bacterial system. While all the N-terminal SAK fusions were found to self-cleave in crude and purified preparations, the C-terminal SAK fusion was stable. The cleavage property of Staphylokinase fusion proteins, inhibited by reduced glutathione and PMSF, was independent of its thrombolytic activity and also independent on the type of host employed for its expression. The serine protease domain of the SAK gene possibly lies between 20th to 77th amino acid and serine 41 of this region appears critical for such a cleavage property.
authors.affiliation: Biotechnology R&D, Lupin Limited, Gat # 1156, Ghotawade Village, Mulshi Taluka, Pune 411042, India.
authors.forename: Bhaskarjyoti
authors.initials: B
authors.lastname: Prasad
authors.forename: Shardul S
authors.initials: SS
authors.lastname: Salunkhe
authors.forename: Sriram
authors.initials: S
authors.lastname: Padmanabhan
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000201:enzymology
chemical_list: D011993:Recombinant Fusion Proteins; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D005978:Glutathione