Characterization of a recombinant chimeric plasminogen activator composed of Gly-Pro-Arg-Pro tetrapeptide and truncated urokinase-type plasminogen activator expressed in Escherichia coli.
A chimeric plasminogen activator, GPRP-u-PA (144-411), consisting of the Gly-Pro-Arg-Pro tetrapeptide fused to the N-terminal of a truncated urokinase-type plasminogen activator (comprising Leu 144 through Leu 411), was produced by expression of the corresponding chimeric cDNA in Escherichia coli cells. After renaturation, the chimera was purified to homogeneity with specific amidolytic activity of 100,000 IU/mg protein. The chimera showed 6-fold greater affinity for fibrin clots than native low molecular weight urokinase (LUK) and 1.5-fold greater affinity than a chemical conjugate, GPRP-LUK, generating via coupling Gly-Pro-Arg-Pro tetrapeptide to native low molecular weight urokinase. The chimera had 2 to 3 fold greater fibrinolytic potency than native LUK in vitro. Fibrinogen had no influence on fibrinolysis of the chimera. The chimera consumed much less fibrinogen than native LUK.
Hua
Z C
ZC
Department of Biochemistry, Nanjing University, People's Republic of China.
Chen
X C
XC
Dong
C
C
Zhu
D X
DX
D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism / Q000494:pharmacology; D005342:Fibrinolysis; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides / Q000096:biosynthesis / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*
D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C034585:glycyl-prolyl-arginyl-proline; D005337:Fibrin; D005340:Fibrinogen; D014568:Urokinase-Type Plasminogen Activator