Design of a novel plasminogen activator based on the structure of hirudin.
Using a phage library, seven peptide sequences with high affinity to human microplasminogen were obtained. Caseinolytic assay indicated that only the synthesized peptide P07 had slight fibrinolytic activity. To enhance its plasminogen activation ability, peptide P07 was fused into loop 32-35 of hirudin. In vitro assay demonstrated that this hirudin-like fusion protein can activate human plasminogen and retain the function of thrombin inhibition. Fusing the sequence ''SPDASRL'' into hirudin generated a plasminogen activation activity 100 times higher than peptide P07 in chromogenic and radial caseinolytic assay. This significant functional improvement might originate from a more specific active structure due to the hirudin scaffold.
Zhang
Yu-Gao
YG
The Key Laboratory of Molecular Medicine, Ministry of Education, Fudan University, Shanghai 200032, China.
Yue
Lu
L
Wang
Yu-Xiong
YX
Tao
Xian-Mei
XM
Song
Hou-Yan
HY
D000595:Amino Acid Sequence; D002364:Caseins / Q000378:metabolism; D015195:Drug Design; D006629:Hirudins / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D010960:Plasminogen Activators / Q000737:chemistry* / Q000494:pharmacology*; D012204:Rh-Hr Blood-Group System / Q000737:chemistry* / Q000494:pharmacology*; D016415:Sequence Alignment; D013917:Thrombin / Q000037:antagonists & inhibitors
D002364:Caseins; D006629:Hirudins; D019151:Peptide Library; D010455:Peptides; C066468:RHCE protein, human; D012204:Rh-Hr Blood-Group System; D010960:Plasminogen Activators; D013917:Thrombin