Kinetic studies on novel plasminogen activators. Demonstration of fibrin enhancement for hybrid enzymes comprising the A-chain of plasmin (Lys-78) and B-chain of tissue-type plasminogen activator (Ile-276) or urokinase (Ile-159).
The activation of plasminogen by two novel hybrid enzymes, constructed from the A-chain of plasmin and the B-chains of tissue-type plasminogen activator (t-PA) or urokinase, was compared with the activation by the parent enzymes. Basal kinetic constants for 'Lys-plasminogen' (human plasminogen with N-terminal lysine) and 'Glu-plasminogen' (human plasminogen with N-terminal glutamic acid) activation were similar to those of the parent activators. The Km for plasminogen turnover for both hybrid enzymes was considerably decreased in the presence of both soluble fibrin and a mimic, a CNBr digest of fibrinogen. These enhancements and the related apparent negative co-operativity are similar to the behaviour of t-PA itself. The results are discussed with regard to the molecular features involved in the mechanism of fibrin stimulation.
Fears
R
R
Beecham Pharmaceuticals Research Division, Epsom, Surrey, U.K.
Dodd
I
I
Ferres
H
H
Robinson
J H
JH
D015119:Aminocaproic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005337:Fibrin / Q000494:pharmacology*; D005341:Fibrinolysin / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D010446:Peptide Fragments / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism*; D010960:Plasminogen Activators / Q000493:pharmacokinetics*; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000494:pharmacology*
D010446:Peptide Fragments; C018998:lysyl-plasminogen; D005337:Fibrin; D010958:Plasminogen; D010960:Plasminogen Activators; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; D015119:Aminocaproic Acid