Purification of gamma-glutamyltransferases by immunoaffinity chromatography and some properties of the enzymes.
Immunoadsorbents were obtained by coupling antibodies to Sepharose 4 B activated with cyanogen bromide. Thus immobilized antibody directed against bovine kidney gamma-glutamyltransferase was used for immunoaffinity chromatography of the enzyme from bovine kidney and liver, from cow milk and from sheep kidney and liver. Immobilized anti-rat kidney gamma-glutamyltransferase antibody was used for purification of the enzyme from rat kidney, mouse kidney, hamster kidney and rat Morris hepatoma 5123D. Yields of the protease-solubilized gamma-glutamyltransferases isolated on immunoadsorbents columns were usually over 60%. The purified enzymes were almost homogenous in polyacrylamide gel electrophoresis. The enzymes showed different molecular weights and electrophoretic mobilities. The effect of antibodies on affinity of the enzymes to substrate and inhibition by synthetic anthglutin and its isomers were studied.
Prusak
E
E
Szewczuk
A
A
D000818:Animals; D000906:Antibodies; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D005260:Female; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008114:Liver Neoplasms, Experimental / Q000201:enzymology; D051379:Mice; D008892:Milk / Q000201:enzymology; D008970:Molecular Weight; D009928:Organ Specificity; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D005723:gamma-Glutamyltransferase / Q000302:isolation & purification* / Q000378:metabolism
D000906:Antibodies; D005723:gamma-Glutamyltransferase