Construction and characterization of a novel staphylokinase variant with thrombin-inhibitory activity.
Staphylokinase (SAK) is an effective thrombolysis agent for therapy of myocardial infarction. We have constructed a fusion SAK variant (SAK-HV) with a thrombin-binding domain composed of 12 amino acids from hirudin and expressed it in Escherichia coli and purified the resultant protein. SAK-HV maintained fibrinolytic activity similar to SAK and had anticoagulant activity attributable to its hirudin segment. Measurement of thrombin-binding activity in vitro demonstrated that SAK-HV possessed binding activity with thrombin while SAK did not. SAK-HV might thus be a more potent thrombolytic agent with anticoagulation property than SAK.
Wang
Min
M
Laboratory of Molecular Genetics, Beijing Institute of Basic Medical Sciences, Beijing, 100850, China.
Wang
Yuanyuan
Y
Wang
Jinfeng
J
Zou
Minji
M
Liu
Shen
S
Xu
Tao
T
Cai
Xin
X
Wu
Chen
C
Wang
Jiaxi
J
Xu
Donggang
D
D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006629:Hirudins / Q000235:genetics* / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013917:Thrombin / Q000037:antagonists & inhibitors*
D006629:Hirudins; D011993:Recombinant Fusion Proteins; D013917:Thrombin; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus