Neurochemical Research

, Volume 10, Issue 10, pp 1325–1334 | Cite as

Partial purification of γ-glutamyltransferase from human brain microvessels

  • Jaroslav Veselý
  • Václav Lisý
  • Miroslav Černoch
Original Articles

Abstract

Two forms of gamma-glutamyltransferase from human brain cortex microvessels were partially purified by gel permeation and ion-exchange and group-affinity chromatography. The specific activity of the purified preparations was 320-fold (detergent form) and 830-fold (proteolytic form) higher than that of the enzyme in the brain cortex homogenate. The relative molecular mass of the proteolytic form of the enzyme was about 90,000 as determined by gel permeation chromatography. The major part of the enzyme (about 80%) was absorbed on Con A-Sepharose 4B. The pH optima for transfer reactions with γ-glutamyl-4-nitroanilide as donor and glycylglycine andl-cystine as acceptors were in the range of 8.2 to 9.0. The studied enzyme was inhibited by a mixture ofl-serine and borate and by bromcresol green.

Keywords

Human Brain Borate Brain Cortex Partial Purification Relative Molecular Mass 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Curthoys, N. P., Hughey, R. P. 1979. Characterization and physiological function of rat renal γ-glutamyltranspeptidase. Enzyme 24:583–403.Google Scholar
  2. 2.
    Meister, A. 1981. On the cycles of glutathione metabolism and transport. Pages 21–58,in Estabrook, R. W., andSrere, P., (eds.), Current Topics in Cellular Regulation, Vol. 18, Biological Cycles, Academic Press, New York.Google Scholar
  3. 3.
    Zelazo, P., andOrlowski, M. 1976. γ-Glutamyl transpeptidase of sheep-kidney cortex. Isolation, catalytic properties and dissociation into two polypeptide chains. Eur. J. Biochem. 61:147–155.PubMedGoogle Scholar
  4. 4.
    Tate, S. S., andMeister, A. 1975. Identity of maleate-stimulated glutaminase with γ-glutamyl transpeptidase in rat kidney. J. Biol. Chem. 250:4619–4627.PubMedGoogle Scholar
  5. 5.
    Hughey, R. P., andCurthoys, N. P. 1976. Comparison of the size and physical properties of γ-glutamyltranspeptidase purified from rat kidney following solubilization with papain or with Triton X-100. J. Biol. Chem. 251:7863–7870.Google Scholar
  6. 6.
    Tate, S. S., andRoss, M. E. 1977. Human kidney γ-glutamyl transpeptidase; catalytic properties, subunits structure, and localization of the γ-glutamyl binding site on the light subunit. J. Biol. Chem. 252:6042–6045.PubMedGoogle Scholar
  7. 7.
    Takahashi, S., Zukin, R. S., andSteinman, H. M. 1981. γ-Glutamyl transpeptidase from WI-38 fibroblasts: purification and active site modification studies. Arch. Biochem. Biophys. 207:87–95.PubMedGoogle Scholar
  8. 8.
    Yokosawa, N., Taniguchi, N., Tsukada, Y., andMakita, A. 1981. Physicochemical and immunochemical characterization of γ-glutamyl transpeptidase from yolk sac tumor and ascitic hepatoma (AH-66) cells. Oncodev. Biol. Med. 2:165–177.PubMedGoogle Scholar
  9. 9.
    Hata, K., Hayakawa, M., Abiko, Y., andTakiguchi, H. 1981. Purification and properties of γ-glutamyl transpeptidase from bovine parotid gland. Int. J. Biochem. 13:681–692.PubMedGoogle Scholar
  10. 10.
    Furukawa, M., Higashi, T., Tateishi, N., Ochi, K. andSakamoto, Y. 1983. Purification and properties of bovine liver γ-glutamyltransferase. J. Biochem. 93:839–846.PubMedGoogle Scholar
  11. 11.
    Horiuchi, S., Inoue, M., andMorino, Y. 1978. γ-Glutamyl transpeptidase: sidedness of its active site on renal brush border membrane. Eur. J. Biochem. 97:429–437.Google Scholar
  12. 12.
    Tsuji, A., Matsuda, Y., andKatunuma, N. 1980. Studies on the structure of γ-glutamyltranspeptidase. II. Location of the segment anchoring γ-glutamyltranspeptidase to the membrane. J. Biochem. 87:1567–1571.PubMedGoogle Scholar
  13. 13.
    Albert, Z., Orlowski, M., Rzucidlo, Z., andOrlowska, J. 1966. Studies on γ-glutamyl transpeptidase activity and its histochemical localization in the central nervous system of man and different animal species. Acta Histochem. 25:312–320.PubMedGoogle Scholar
  14. 14.
    Meister, A., Tate, S. S., andRoss, L. L. 1976. Membrane-bound γ-glutamyl transpeptidase. Pages 315–347,in Martonosi, A. (ed.), The Enzymes of Biological Membranes, Vol. 3, Plenum Press, New York.Google Scholar
  15. 15.
    Shine, H. D., andHaber, B. 1981. Immunocytochemical localization of γ-glutamyl transpeptidase in the rat CNS. Brain Res. 217:339–349.PubMedGoogle Scholar
  16. 16.
    Reyes, E., andBarela, T. D. 1980. Isolation and purification of multiple forms of γ-glutamyl transpeptidase from rat brain. Neurochem. Res. 5:159–170.Google Scholar
  17. 17.
    Tate, S. S., Ross, L. L., andMeister, A. 1973. The γ-glutamyl cycle in the choroid plexus: its possible function in amino acid transport. Proc. Natl. Acad. Sci. USA 70:1447–1449.PubMedGoogle Scholar
  18. 18.
    Prusiner, P. E., andPrusiner, S. B. 1978. Partial purification and kinetics of γ-glutamyl transpeptidase from bovine choroid plexus. J. Neurochem. 30:1253–1259.PubMedGoogle Scholar
  19. 19.
    Lisý, V., Šťastný, F., andLodin, Z. 1979. Regional distribution of membrane-bound γ-glutamyl transpeptidase activity in mouse brain. Comparison with rabbit brain. Neurochem. Res. 4:747–753.Google Scholar
  20. 20.
    Grandgeorge, M., andMorélis, P. 1976. Purification partielle et étude de la γ-glutamyl transpeptidase des capillaires cérébraux de mouton. Biochimie 58:275–284.PubMedGoogle Scholar
  21. 21.
    Veselý, J., andČernoch, M. 1984. Studies of γ-glutamyltransferase activity in the brain tissue post mortem. Neurochem. Res. 9:917–925.PubMedGoogle Scholar
  22. 22.
    Veselý, J., andČernoch, M. 1984. Solubilization and some properties of γ-glutamyltransferase from human brain microvessels. Neurochem. Res. 9:927–934.PubMedGoogle Scholar
  23. 23.
    Kulhánek, V., andDimov, D. M. 1973. Bestimmung der γ-Glutamyltranspeptidase. Ergebnisse Exp. Med. 12:161–169.Google Scholar
  24. 24.
    Lowry, O. H., Rosenbrough, N. R., Farr, A. L. L., andRandall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275.PubMedGoogle Scholar
  25. 25.
    Brendel, K., Meezan, E., andCarlson, E. C. 1974. Isolated brain microvessles: a purified, metabolically active preparation from bovine cerebral cortex. Science 185:953–955.PubMedGoogle Scholar
  26. 26.
    Shaw, L. M., London, J. W., andPetersen, L. E. 1978. Isolation of γ-glutamyltransferase from human liver, and comparison with the enzyme from human kidney. Clin. Chem. 24:905–915.PubMedGoogle Scholar
  27. 27.
    Huseby, N.-E. 1977. Purification and some properties of γ-glutamyltransferase from human liver. Biochim. Biophys. Acta 483:46–56.PubMedGoogle Scholar
  28. 28.
    Thompson, G. A., andMeister, A. 1975. Utilization of L-cystine by the γ-glutamyl transpeptidase-γ-glutamyl cyclotransferase pathway. Proc. Natl. Acad. Sci. USA 72:1985–1988.PubMedGoogle Scholar
  29. 29.
    Binkley, F. 1961. Purification and properties of renal glutathionase. J. Biol. Chem. 236:1075–1082.Google Scholar
  30. 30.
    Kozak, E. M., andTate, S. S. 1980. Interaction of the antitumor drug,l-(αS,5S)-α-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid (AT-125) with renal brush border membranes. Specific labeling of γ-glutamyl transpeptidase. FEBS Lett. 122:175–178.PubMedGoogle Scholar
  31. 31.
    Köttgen, E., andGerok, W. 1976. Isoenzymdifferenzierung der Gamma-Glutamyltransferase mit Concanavalin A und Con A-Sepharose. Klin. Wochschr. 54:439–444.Google Scholar
  32. 32.
    Scherberich, J. E., Kleemann, B., andMondorf, W. 1979. Isolation of kidney brush border gamma-glutamyl transpeptidase from urine by specific antibody gel chromatography. Clin. Chim. Acta 93:35–41.PubMedGoogle Scholar
  33. 33.
    Huseby, N.-E. 1981. Separation and characterization of human γ-glutamyltransferases. Clin. Chim. Acta 111:39–45.PubMedGoogle Scholar
  34. 34.
    Yamashita, K., Hitoi, A., Tateishi, N., Higashi, T., Sakamoto, Y., andKobata, A. 1983. Organ-specific difference in the sugar chains of γ-glutamyltranspeptidase. Arch. Biochem. Biophys. 225:993–998.PubMedGoogle Scholar
  35. 35.
    Tate, S. S., andMeister, A. 1978. Serine-borate complex as a transition-state inhibitor of γ-glutamyl transpeptidase. Proc. Natl. Acad. Sci. USA 75:4806–4809.PubMedGoogle Scholar
  36. 36.
    Thompson, G. A., andMeister, A. 1977. Interrelationships between the binding sites for amino acids, dipeptides, and γ-glutamyl donors in γ-glutamyl transpeptidase. J. Biol. Chem. 252:6792–6798.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1985

Authors and Affiliations

  • Jaroslav Veselý
    • 1
  • Václav Lisý
    • 2
  • Miroslav Černoch
    • 1
  1. 1.Institute of ChemistryPalacký University Medical FacultyOlomoucCzechoslovakia
  2. 2.Institute of PhysiologyCzechoslovak Academy of SciencesPragueCzechoslovakia

Personalised recommendations