Alpha-Aminoadipate aminotransferase and kynurenine aminotransferase. Purification, characterization, and further evidence for identity.
Alpha-Aminoadipate aminotransferase and kynurenine aminotransferase activities were co-purified from the rat kidney supernatant fraction. The resulting preparation was determined to be nearly homogenous by analytical disc gel electrophoresis at pH 8.9 and 7.5 isoelectric focusing on polyacrylamide gels, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A molecular weight of approximately 85,000 was determined on Sephadex G-200 chromatography and sucrose density gradient analysis. The enzyme was determined to be comprised of two subunits of approximately the same molecular weight (45,500 +/- 850) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. An isoelectric pH of 6.56 +/- 0.06 was determined by focusing on polyacrylamide gels. Further evidence is provided to support the idea that the alpha-aminoadipate aminotransferase and kynureine aminotransferase activities are properties of a single protein: (a) co-purification of the two activities from the rat kidney supernatant fraction with the ratio of their specific activities remaining constant, (b) similar chromatographic behavior, (c) a similarity in their dependence on added pyridoxal-P for activity, and (d) a similar pattern of heat inactivation.
D015074:2-Aminoadipic Acid; D000818:Animals; D001219:Aspartate Aminotransferases / Q000302:isolation & purification; D055598:Chemical Phenomena; D002621:Chemistry; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004589:Electrophoresis, Disc; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006882:Hydroxyapatites; D066298:In Vitro Techniques; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology / Q000648:ultrastructure; D007737:Kynurenine; D008970:Molecular Weight; D051381:Rats; D012694:Serine / Q000494:pharmacology; D000637:Transaminases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism
D006882:Hydroxyapatites; D015074:2-Aminoadipic Acid; D007737:Kynurenine; D012694:Serine; D000637:Transaminases; D001219:Aspartate Aminotransferases