Purification and characterization of kynurenine-pyruvate aminotransferase from rat kidney and brain.
Kynurenine-pyruvate aminotransferase (KPT), the enzyme responsible for the biosynthesis of the endogenous excitatory amino acid receptor antagonist kynurenic acid, was purified to homogeneity from rat kidney, as judged by polyacrylamide and sodium dodecyl sulfate electrophoresis. The protein appeared to consist of 2 identical subunits of approximately 48 kDa. Kinetic analysis showed Km values of 2.8 mM (kynurenine) and 3.8 mM (pyruvate), respectively. KPT was also partially purified from rat brain. Kidney and brain KPT were found to be identical when analyzed by a spectrum of biochemical, physico-chemical and, after production of anti-kidney KPT antibodies, immunological methods. Partially purified anti-KPT antiserum was used for first immunohistochemical studies, which revealed the presence of the enzyme in astrocyte-like cells throughout the brain. Less frequently, KPT was also found in discretely arranged neurons. The availability of pure KPT and specific anti-KPT antibodies can be expected to be of value for the further examination of the neurobiology of kynurenic acid.
Okuno
E
E
Wakayama Medical College, Department of Biochemistry, Japan.
Du
F
F
Ishikawa
T
T
Tsujimoto
M
M
Nakamura
M
M
Schwarcz
R
R
Kido
R
R
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D002845:Chromatography; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D017886:Durapatite; D006651:Histocytochemistry; D006882:Hydroxyapatites; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008190:Lyases; D008970:Molecular Weight; D011773:Pyruvates / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D000637:Transaminases / Q000302:isolation & purification* / Q000378:metabolism
D006882:Hydroxyapatites; D011773:Pyruvates; D017886:Durapatite; D000637:Transaminases; C427605:glutamine - phenylpyruvate transaminase; C021481:kynurenine-oxoglutarate transaminase; D008190:Lyases