Purification and properties of gamma-glutamyl transpeptidase from human testis.
Gamma-glutamyl transpeptidase was purified to apparent homogeneity from human testis by DEAE cellulose, acetone, precipitation, fractionation by ammonium sulphate, Sephacryl S-200 chromatography, Q-Sepharose chromatography and S-Sepharose chromatography following solubilization of the enzyme by Triton X-100. The purified enzyme had an apparent molecular weight of 54 KDa by Sephacryl S-200 gel filtration. On sodium dodecyl sulphate polyacrylamide gel electrophoresis, two submits of molecular weight 38 KDa and 14 KDa were obtained. The purified enzyme showed a single band with pI 6.0. The Km value and the optimal pH of the enzyme for L-gamma-glutamyl-3-carboxy-4-nitroanilide were found 1.09 mmol/l and 8.2-8.5, respectively. Serial lectin binding study with various lectin columns showed that the majority of the asparagine-linked oligosaccharides of the enzyme was complex-types. However, complex-types with bisecting N-acetylglucosamine residue were not recognized.
Yoshida
K
K
Department of Urology, Saitama Medical Center, Medical School, Japan.
Arai
K
K
Kobayashi
N
N
Saitoh
H
H
D000368:Aged; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005590:Fractional Precipitation; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D037102:Lectins / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D013737:Testis / Q000201:enzymology*; D005723:gamma-Glutamyltransferase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D037102:Lectins; D005723:gamma-Glutamyltransferase