Mitochondrial aspartate aminotransferase: a third kynurenate-producing enzyme in the mammalian brain.
The tryptophan metabolite kynurenic acid (KYNA), which is produced enzymatically by the irreversible transamination of l-kynurenine, is an antagonist of alpha7 nicotinic and NMDA receptors and may thus modulate cholinergic and glutamatergic neurotransmission. Two kynurenine aminotransferases (KAT I and II) are currently considered the major biosynthetic enzymes of KYNA in the brain. In this study, we report the existence of a third enzyme displaying KAT activity in the mammalian brain. The novel KAT had a pH optimum of 8.0 and a low capacity to transaminate glutamine or alpha-aminoadipate (the classic substrates of KAT I and KAT II, respectively). The enzyme was inhibited by aspartate, glutamate, and quisqualate but was insensitive to blockade by glutamine or anti-KAT II antibodies. After purification to homogeneity, the protein was sequenced and the enzyme was identified as mitochondrial aspartate aminotransferase (mitAAT). Finally, the relative contributions of KAT I, KAT II, and mitAAT to total KAT activity were determined in mouse, rat, and human brain at physiological pH using anti-mitAAT antibodies. KAT II was most abundant in rat and human brain, while mitAAT played the major role in mouse brain. It remains to be seen if mitAAT participates in cerebral KYNA synthesis under physiological and/or pathological conditions in vivo.
Guidetti
Paolo
P
Maryland Psychiatric Research Center, University of Maryland School of Medicine, Baltimore, Maryland 21228, USA.
Amori
Laura
L
Sapko
Michael T
MT
Okuno
Etsuo
E
Schwarcz
Robert
R
D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism*; D001921:Brain / Q000201:enzymology*; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D015723:Gene Library; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007120:Immunochemistry; D007527:Isoenzymes / Q000378:metabolism; D007736:Kynurenic Acid / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D008190:Lyases / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000201:enzymology*; D016318:Quisqualic Acid / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D000637:Transaminases / Q000378:metabolism
D018076:DNA, Complementary; D018690:Excitatory Amino Acid Agonists; D007527:Isoenzymes; D016318:Quisqualic Acid; D000637:Transaminases; D001219:Aspartate Aminotransferases; C427605:glutamine - phenylpyruvate transaminase; C021481:kynurenine-oxoglutarate transaminase; D008190:Lyases; D007736:Kynurenic Acid