In Vitro Effects of γ-Glutamyltranspeptidase Inhibitor Acivicin on Human Myeloid and B Lineage Cells
Gamma-glutamyltranspeptidase (γ-GT, EC 22.214.171.124) (Figure 1) is a widely distributed enzyme that acts as a transferase in the transfer of the γ-glutamyl group from a wide variety of peptide donors to other amino acids and peptide acceptors, and as a hydrolase in removing the γ-glutamyl residue from such peptides . γ-GT activity was initially detected at the surface of human monocytes . Using a spectrophotometric method, we showed it to be a sensitive assay of γ-GT activity detected on the surface of intact cells, as well as in highly purified cell membrane fractions. Cell-surface γ-GT (assayed with γ—Glu-para-nitroanilide as substrate and Gly-Gly as acceptor and abolished by the specific inhibitor of γ-GT i.e. acivicin) was confirmed in the myeloblastic (HL-60) and monoblastic (U937) cell lines . Peripheral blood monocytes, granulocytes as well as macrophages developed in vitro from monocytes. were also found to exhibit γ-GT activity . Another line of evidence that human myeloid cells express γ-GT was obtained from Northern blot analysis. In humans, there are at least four potential genes for γ-GT located on chromosome 22 . Using a liver γ-GT probe, we have detected an mRNA species of 2.4 kb, corresponding to that previously described for the HepG2 cell line  in both cell lines and in granulocytes and macrophages . Resting human B cells purified from blood as well as plasma cell lines (U266, RPMI 8226, Eskol) expressed γ-GT activity. However, no γ-GT transcripts were detected in any cells of the B lineage.
KeywordsU266 Cell Peripheral Blood Monocyte Vitro Effect Human Myeloid Cell Ectoenzyme Activity
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- 1.Tate SS, Meister A. (1981) y-glutamyl transpeptidase: catalytic, structural and functional aspects. Mot Cell. Biol. 39: 357–368.Google Scholar
- 3.Bauvois B, Laouar A, Rouillard D, Wietzerbin J. (1995) Inhibition of y-glutamyl transpeptidase activity at the surface of human myeloid cells is correlated with macrophage maturation and transforming growth factor-13 production. Cell Growth und Differentiation 6: 1163–1170.Google Scholar
- 4.Guellaën C. (1989) Röle physiologique de la y-glutamyl transpeptidase. Med. Sci. 5: 637–644.Google Scholar
- 8.Shipp M, Look AT. (1993) Hematopoietic differentiation antigens that are membrane-associated enzymes: cutting is the key. Blood 68: 1052–1070.Google Scholar
- 1.Bond, J. S., and A. J. Barrett, 1993, Proteolysis and protein turnover, Proceeding of the 9th I(.’OP Meeting. Williamsburg, Virginia, U. S. A., Portland Press Proceedings. page X IV.Google Scholar
- 2.Hoesch, Kurt, 1921, Emil Fischer. sein Leben und sein Werk, Verlag Chemie, G.m.b.11.. Berlin und Leipzig, 480 pages.Google Scholar
- 3.Abderhalden. Emil. 1906. Lehrbuch der Physiologischen Chemie, Urbahn und Schwarzenberg, Berlin. Wien, 787 pages.Google Scholar