Partial purification of γ-glutamyltransferase from human brain microvessels
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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.
KeywordsHuman Brain Borate Brain Cortex Partial Purification Relative Molecular Mass
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- 1.Curthoys, N. P., Hughey, R. P. 1979. Characterization and physiological function of rat renal γ-glutamyltranspeptidase. Enzyme 24:583–403.Google Scholar
- 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
- 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
- 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
- 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
- 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
- 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
- 23.Kulhánek, V., andDimov, D. M. 1973. Bestimmung der γ-Glutamyltranspeptidase. Ergebnisse Exp. Med. 12:161–169.Google Scholar
- 29.Binkley, F. 1961. Purification and properties of renal glutathionase. J. Biol. Chem. 236:1075–1082.Google Scholar
- 31.Köttgen, E., andGerok, W. 1976. Isoenzymdifferenzierung der Gamma-Glutamyltransferase mit Concanavalin A und Con A-Sepharose. Klin. Wochschr. 54:439–444.Google Scholar