Cloning and recombinant expression of rat and human kynureninase

FEBS Letters
S TomaL Benatti

Abstract

Kynureninase [E.C.3.7.1.3.] is one of the enzymes involved in the biosynthesis of NAD cofactors from tryptophan through the kynurenine pathway. By tryptic and CNBr digestion of purified rat liver kynureninase, we obtained about 28% of the amino acid sequence of the enzyme. The rat kynureninase cDNA, isolated by means of reverse-transcribed polymerase chain reaction and hybridization screening, codes for a polypeptide of 464 amino acids. Northern blot analysis revealed the synthesis of a 2.0 kb rat kynureninase mRNA. A cDNA encoding human liver kynureninase was also isolated. The deduced amino acid sequence is 85% identical to that of the rat protein. COS-1 cells were transfected with both cDNAs. The Km values of the rat enzyme, for L-kynurenine and DL-3-hydroxykynurenine, were 440 +/- 20 microM and 32 +/- 5 microM and of the human enzyme 440 /- 20 microM and 49 +/- 6 microM, respectively. Interestingly, COS-1 cells transfected with the cDNA coding for rat kynureninase also display cysteine-conjugate beta-lyase activity.

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Citations

Mar 6, 2007·Journal of Inherited Metabolic Disease·M ChristensenE Christensen
Feb 16, 2007·Biochemistry·Santiago LimaRobert S Phillips
May 3, 2002·European Journal of Biochemistry·Harold A Walsh, Nigel P Botting
Sep 14, 2011·Nucleic Acids Research·Vincenzo CalvaneseMario F Fraga
Apr 2, 2002·Hypertension Research : Official Journal of the Japanese Society of Hypertension·Ken-ichi MizutaniNaoharu Iwai
Jan 28, 2009·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Yan MengToru Shimada
Nov 24, 2020·Future Virology·Rafeed Rahman TurjyaAbul Bashar Mir Md Khademul Islam
Apr 9, 2021·Pharmacology & Therapeutics·Marta Marszalek-GrabskaWaldemar A Turski

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