A rapid, high-yield purification of L-alanine:4,5-dioxovalerate transaminase from rat kidney mitochondria using an improved enzyme assay method.
The present report documents an improved enzyme assay method for the mammalian L-alanine:4,5-dioxovalerate transaminase which is of significant utility in work with crude tissue homogenates, cell cultures, or purified enzyme preparations. We also describe a new and rapid purification procedure for this enzyme from rat kidney mitochondria. The three-step procedure involves the use of digitonin and lubrol for mitochondrial matrix preparation and L-alanine-Sepharose 4B column chromatography followed by gel filtration on Sepharose 6B. By this procedure it is possible to obtain a highly purified enzyme preparation in a relatively short time with a 37.5% yield.
Tyagi
R K
RK
Biochemistry Laboratory, School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India.
Datta
K
K
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003429:Cross Reactions; D004072:Digitonin; D007668:Kidney / Q000201:enzymology*; D008928:Mitochondria / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D011092:Polyethylene Glycols; D011233:Precipitin Tests; D051381:Rats; D000637:Transaminases / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*
D011092:Polyethylene Glycols; C005092:lubrol; D000637:Transaminases; C019687:aminolevulinate aminotransferase; D004072:Digitonin