Hydrolysis of L-Histidine Methyl Ester

Summary

The ratio of TAME esterase activity (or PME esterase activity for α-chymotrypsin) to HME esterase activity has been determined for a number of proteolytic enzymes under identical conditions. Papain showed no attack on HME, but ficin, bromelin, human plasmin, trypsin, thrombin, autoprothrombin C, urokinase and α-chymotrypsin hydrolyze this substrate to varying degrees. Ficin showed a TAME/HME activity ratio of 304 while the lowest ratio (approximately 1) was found for urokinase, an activator of plasminogen. This high relative affinity of urokinase for HME prompted an investigation to determine if this property was also a function of streptokinase-activator complex. Glycerol-activated plasmin had a low relative affinity for HME as a substrate (TAME/HME = 157), but addition of high concentrations of streptokinase increased the relative affinity of the preparation for HME catalysis by 500 to 600% (TAME/HME = 26.4). Human plasminogen activated by high concentrations of streptokinase also demonstrate high relative affinity for HME (TAME/ HME = 20-25). HME esterase activity is largely lost when streptokinase is destroyed by pH 2.0 treatment, but can be regained by readdition of streptokinase to the plasmin preparation. Hydrolysis of HME by plasmin preparations in the presence of streptokinase is believed to be primarily a property of streptokinase-plasmin complex.

^* This investigation was supported by the National Heart Institute, National Institutes of Health, under Grant No. HE-10146-02. Some of this material was presented at the Fifteenth Annual Symposium on Blood, Wayne State University School of Medicine, Detroit, Michigan, January 20th and 21st, 1967.

• References

• 1 Bergmann M, Fruton J.S. The specificity of proteinases. Adv. in Enzymol. 1941; 1: 63
• 2 Cole E.R. Hydrolysis of L-histidine methyl ester. I. The action of thrombin. Thrombos. Diathes. haemorrh. (Stuttg.) 1968; 19: 321
• 3 Cole E.R, Koppel J.L, Olwin J.H. The multiple specificity of thrombin for synthetic substrates. Nature 1967; 213: 405
• 4 Bergmann M, Fruton J.S, Pollok H. The specificity of trypsin. J. Biol. Chem. 1939; 127: 643
• 5 Neurath H, Schwert G.W. The mode of action of the crystalline pancreatic proteolytic enzymes. Chem. Rev. 1950; 46: 69
• 6 Neil G.L, Niemann G, Hein G.E. Structural specificity of A-chymotrypsin: polypeptide substrates. Nature 1966; 210: 903
• 7 Seegers W.H. The purification of prothrombin. Record Chem. Progr. (Kresge-Hooker Sci. Lib.) 1952; 13: 143
• 8 Seegers W.H, Cole E.R, Harmison G.R, Marciniak E. Purification and some properties of autoprothrombin C. Canad. J. Biochem. Physiol. 1963; 41: 1047
• 9 Seegers W.H, Smith H.P. Factors which influence the activity of purified thrombin. Amer. J. Physiol. 1942; 137: 348
• 10 Cole E.R, Marciniak E, Seegers W.H. Procedures for the quantitative determination of autoprothrombin C. Thrombos. Diathes. haemorrh. (Stuttg.) 1962; 8: 434
• 11 Kline D.L. The purification and crystallization of plasminogen (Profibrinolysin). J. Biol. Chem. 1953; 204: 949
• 12 Roberts P.S. Measurement of the rate of plasmin action on synthetic substrates. J. Biol. Chem. 1958; 252: 285
• 13 Roberts P.S. The esterase activities of human plasmin during purification and subsequent activation by streptokinase or glycerol. J. Biol. Chem. 1960; 235: 2262
• 14 Remmert L.F, Cohen P. Partial purification and properties of a proteolytic enzyme of human serum. J. Biol. Chem. 1949; 181: 431
• 15 Kline D.L, Fishman J.B. Proactivator function of human plasmin as shown by lysine esterase assay. J. Biol. Chem. 1961; 236: 2807
• 16 Alkjaersig N, Fletcher A.P, Sherry S. Activation of human plasminogen. II. A kinetic study of activation with trypsin, urokinase and streptokinase. J. Biol. Chem. 1958; 233: 86
• 17 Engel A, Alexander B, Pechet L. Activation of trypsinogen and plasminogen by thrombin preparations. Biochemistry 1966; 5: 1543
• 18 Sherry S, Alkjaersig N, Fletcher A.P. Activity of plasmin and streptokinase-activator on substituted arginine and lysine esters. Thrombos. Diathes. haemorrh. (Stuttg.) 1966; 16: 18
• 19 Erlanger B.F, Castleman H. On the activation of trypsin by amines. Biochim. Biophys. Acta 1964; 85: 507