Comparison of fibrin-mediated stimulation of plasminogen activation by tissue-type plasminogen activator (t-PA) and fibrin-dependent enhancement of amidolytic activity of t-PA.
Studies in the past 10 years have shown that there are two different, but related pathways for the acceleration of tissue-type plasminogen activator (t-PA) catalysis: (1) fibrin-dependent enhancement of t-PA amidolytic activity by fibrin binding; (2) fibrin-mediated stimulation of plasminogen activation by t-PA via the formation of a ternary complex of fibrin, t-PA and plasminogen. The common characteristic of both phenomena is the affinity of t-PA for fibrin, which is realized by the same enzyme binding site. However, a comparison of the kinetic data, the participating fibrin structures and the differences between single-chain and two-chain t-PA (sct-PA and tct-PA, respectively) shows that both phenomena have different causes. Fibrin-mediated stimulation of plasminogen activation involves both sct-PA and tct-PA and different fibrinogen derivatives such as fibrin, fibrinogen cyanogen bromide fragment FCB-2, fibrin alpha-chain and poly-lysine. This mechanism is described by a marked apparent decrease in the KM value. In contrast, fibrin-dependent enhancement of t-PA activity against low molecular weight peptides is exclusive to sct-PA and is characterized by an increase in the kcat value and, depending on the nature of the substrate, by an increase in kcat and a decrease in KM. Thus, sct-PA activity modulation depends strictly on the correct three-dimensional folding of fibrin and is not mediated by fibrinogen fragment FCB-2 or isolated fibrin chains.(ABSTRACT TRUNCATED AT 250 WORDS)
Fischer
B E
BE
AFRC Institute of Food Research, Department of Protein Engineering, Reading, UK.
D000577:Amides / Q000097:blood*; D000595:Amino Acid Sequence; D005337:Fibrin / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D010959:Tissue Plasminogen Activator / Q000097:blood*
D000577:Amides; D005337:Fibrin; D010959:Tissue Plasminogen Activator