S-Hippuryl Thioglycolyl Glycine : A New Chromogenic Substrate for Dipeptidyl Carboxypeptidases

Dipeptidyl Carboxypeptidases에 의한 S-Hippuryl Thioglycolyl Glycine의 가수분해

  • Published : 1975.08.30

Abstract

A spectrophotometric assay technique is descriead for the measurement of free SH-groups in the enzyme reaction mixture. The method utilizes a new substrate, S-hippuryl-thioglycolyl-glycine(S-Hip-thioglycol-Gly) which is the basis for a convenient assay of angiotensin-converting enzyme and other dipeptidyl carboxypeptidases. This substrate contains an appropriately located thioester linkage that is hydrolyzed by the converting enzyme and other dipeptidyl carboxypeptidases. One of the products, thioglycolyl glycine, is readily measured by reaction with Ellman's reagent, 5,5'-dithio-bis-(2-nitrobenzoic acid), DTNB, to produce 5-thio-2-nitrobenzoic acid which has a strong absorption band at 410 nm. The method is sensitive (${\varepsilon}M = 1.36{\times}10^4$ at 412 nm) and can be applied as a continuous recording with DTNB present in the enzymatic reaction mixture.

Keywords

References

  1. J. Exptl. Med. v.103 L.T. Skeggs;J.R. Kahn;N.P. Sumway
  2. Nature v.216 K.K.F. Ng.;J.R. Vane
  3. Life Sei. v.2 C.G. Huggins;N.S. Thampi
  4. Acta Path. Microbiol. Scand. v.78 K. Poulsen;J. Bing
  5. J. Pharmacol. Exptl. Therap. v.170 J.D. Battett;M.P. Sambi
  6. Biochim. Biophys Acta v.230 D.W. Cushman;H.S. Cheung
  7. Anal. Biochem. F.E. Dorer;L.T. Skeggs;J.R. Kahn;K.E.Lentz;M. Levine
  8. Biochim. Biophys Acta v.206 Y. Piquilloud;A. Reinharz;M. Roth
  9. Clin. Chim. Acta v.23 H.J.G. Hollemans;J. Jan Dwe Meer;W. Kloosterziel
  10. Biochim. Biophys Acta v.214 H. Y. T. Yang;E. G. Erdos;Y. Levin
  11. J. Pharmacol. Exptl. Therp. v.177 H. Y. T Yang;E. G. Erdos;Y. Levin
  12. Biochem. Pharmacol. v.20 K. W. Cushman;H. S. Cheung
  13. Biochim. Biophys Acta v.276 C. W. Angus;H-J. Lee;I. B. Wilson
  14. Arch. Biochem. Biophys. v.92 G.L. Ellman
  15. Arch. Biochem. Biophys. v.82 G.L. Ellman
  16. Progress in Biochemical Pharmacology v.1 S. Binty-Szlechto;R. Paoletti(ed.);R. Vertua(ed.)
  17. Biochim. Biophys Acta v.309 C. W. Angus;H-J. Lee;I. B. Wilson
  18. Biochim. Biophys Acta v.250 H-J. Lee;J.N. LaRue;I.B. Wilson
  19. Biochim Biophys Acta v.235 H-J. Lee;J.N. LaRueand;I. B. Wilson
  20. Experientia v.25 M. Roth;A.F. Weitzman;Y. Piquilloud