Purification and properties of kynurenine aminotransferase from rat kidney.
Previous reports indicated that a single protein exhibits kynurenine aminotransferase (KAT) and alpha-aminoadipate aminotransferase (AadAT) activities. However, recently we discovered that KAT and AadAT activities are associated with two different proteins. KAT from rat kidney supernatant fraction was purified to electrophoretic homogeneity by (NH4)2SO4 fractionation, DEAE-Sephacel and hydroxyapatite chromatography. This procedure separated KAT from AadAT and improved the overall yield and the degree of purification over previously published methods. Some of the properties of purified KAT, such as Mr, subunit structure and the inhibition by dicarboxylic acids, were identical with those reported previously. However, the substrate specificity and pI of purified KAT were different from earlier reports. The same procedure can also be used to purify KAT from rat kidney mitochondria. These results support our earlier observation that KAT and AadAT activities are associated with two proteins and suggest that cytosolic KAT may be structurally similar to the mitochondrial enzyme.
Mawal
M R
MR
Department of Pediatrics, Children's Hospital of Michigan, Wayne State University, Detroit 48201.
Mukhopadhyay
A
A
Deshmukh
D R
DR
D051307:2-Aminoadipate Transaminase; D000596:Amino Acids / Q000032:analysis; D000645:Ammonium Sulfate; D000818:Animals; D005591:Chemical Fractionation; D002845:Chromatography; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007668:Kidney / Q000201:enzymology* / Q000648:ultrastructure; D007700:Kinetics; D008190:Lyases; D008297:Male; D008928:Mitochondria / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry; D013379:Substrate Specificity; D000637:Transaminases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D000637:Transaminases; D051307:2-Aminoadipate Transaminase; C427605:glutamine - phenylpyruvate transaminase; C021481:kynurenine-oxoglutarate transaminase; D008190:Lyases; D000645:Ammonium Sulfate