Urokinase-like plasma fibrinolytic enzyme induced by Shochu drinking

Abstract

After normal human subjects (78 in number, aged 18-34 years) had taken the local Japanese spirit “Shochu, ” rice wine “Sake” and beer, almost no change in enzyme parameters of plasma fibrinolysis was observed. However, plasma eluate from [N^α-(ε-aminocaproyl)-DL-homoarginine hexylester]-Sepharose affinity chromatography revealed extremely strong pyro-Glu-Gly-Arg-pNA (S-2444) amidolytic and fibrinolytic activities in comparison with those of the control. The highest activities were observed in the Shochu group (1, 133nmol pNA/dl, 470IU/dl plasma), followed by the Sake (811nmol pNA/dl, 415IU/dl plasma) and beer (707nmol pNA/dl, 313IU/dl plasma) groups, corresponding to an approximately 2.1, 1.7 and 1.5 times increase in plasma fibrinolysis compared to the control group (490nmol pNA/dl, 242IU/dl plasma).
Fibrinolytic enzyme isolated from the Shochu group was partially inactivted by rabbit anti HMW-UK serum and could be adsorbed on an anti HMW-UK IgG-Sepharose immunoadsorbent column. The final preparation eluted from the immunoadsorbent column with 6M urea buffer revealed molecular weights of approximately 30, 000 (main band), 50, 000 and 30, 000 (minor bands) by zymography, and an active site of approximately 8% (assuming the molecular weight of 30, 000) by p-nitrophenyl-p′-guanidinobenzoate titration. The enzyme not only hydrolyzed the UK-specific substrate S-2444 but also strongly activated human Glu-plasminogen to plasmin. This indicated that some of the enzyme appearing in the plasma after drinking was “UK-like plasminogen activator”.