Active streptokinase from the cloned gene in Streptococcus sanguis is without the carboxyl-terminal 32 residues.
The streptokinase expressed by the cloned gene in Streptococcus sanguis has a molecular weight of about 44 000 [Malke, H., Gerlach, D., Kohler, W., & Ferretti, J.J. (1984) MGG, Mol. Gen. Genet. 196, 360-365] while the molecular weight of the native streptokinase is 47 000. The structural and activity differences of the cloned streptokinase (cSK) as expressed by S. sanguis and the native streptokinase (nSK) were investigated. From a partially purified cSK, two active fractions were obtained by reversed-phase HPLC. The minor fraction cSKL was nearly as active as SK in plasminogen activation. The major fraction cSKs had only about one-fourth of the specific activity. The structures of cSKL and cSKs were studied and compared to the known amino acid sequence of SK [Jackson, K. W., & Tang, J. (1982) Biochemistry 21, 6620-6625]. From the NH2- and COOH-terminal sequences and amino acid composition of the cyanogen bromide (CNBr) fragments, it could be deduced that cSKL and cSKs are without 31 and 32 residues, respectively, from the COOH-terminal end of SK. Since the cloned gene contained the full SK structure, the missing structures must have been due to posttranslational proteolysis. An SK fragment similar in size to cSK was observed from a chymotryptic digest of SK.
D000596:Amino Acids / Q000032:analysis; D002268:Carboxypeptidases / Q000378:metabolism; D043422:Carboxypeptidases A; D003001:Cloning, Molecular; D003488:Cyanogen Bromide; D005796:Genes; D005798:Genes, Bacterial; D007700:Kinetics; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010958:Plasminogen; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D013298:Streptococcus sanguis / Q000201:enzymology / Q000235:genetics*; D013300:Streptokinase / Q000235:genetics* / Q000378:metabolism
D000596:Amino Acids; D010446:Peptide Fragments; D011994:Recombinant Proteins; D010958:Plasminogen; D002268:Carboxypeptidases; D013300:Streptokinase; D043422:Carboxypeptidases A; D003488:Cyanogen Bromide