Isolation and characterization of a rat liver enzyme with both cysteine conjugate beta-lyase and kynureninase activity.
Cysteine conjugate beta-lyase is a name applied to enzymes which cleave the S-cysteine conjugates of some xenobiotics to pyruvate, ammonia, and a thiol. Recently, several laboratories have characterized these enzymes from kidney, liver, and bacterial sources in an effort to understand their role in the genesis of novel sulfur-containing metabolites of xenobiotics and in the toxicity of some S-cysteine conjugates. Kynureninase is an enzyme which plays a key role in the biosynthesis of nicotinamide ribonucleotides. This investigation demonstrates that rat hepatic cysteine conjugate beta-lyase is the same enzyme as kynureninase. Both activities copurify on ion exchange, hydroxylapatite, and molecular exclusion chromatography. The subunit composition of enzyme prepared by two different methods is identical, Mr = 55,000. The Km values for 3-OH-kynurenine and kynurenine are 13 and 400 microM, respectively. Kynurenine and 3-hydroxykynurenine inhibit cysteine conjugate beta-lyase activity. Inactivation of the enzyme by substrates which undergo beta-elimination results in loss of kynureninase activity, but kynurenine does not inactivate the enzyme. Both enzyme activities react with anti-cysteine conjugate beta-lyase antibody. Product inhibitors of kynureninase, anthranilate, and 3-hydroxyanthranilate are also inhibitors of cysteine conjugate beta-lyase. Heat inactivation at 70 degrees C produced coincident loss of both activities. The enzyme has an absorption maximum at 432 nm suggesting a bound pyridoxal phosphate. These data show that at least one cysteine conjugate beta-lyase is a pyridoxal phosphate enzyme with a biological function other than xenobiotic metabolism. The enzyme can catalyze two distinct types of reactions, i.e. beta-elimination and the kynureninase reaction.
D000818:Animals; D000906:Antibodies; D013437:Carbon-Sulfur Lyases; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006867:Hydrolases / Q000378:metabolism*; D007737:Kynurenine / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008190:Lyases / Q000378:metabolism*; D008297:Male; D011732:Pyridoxal Phosphate / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013056:Spectrophotometry, Ultraviolet
D000906:Antibodies; D007737:Kynurenine; D011732:Pyridoxal Phosphate; D006867:Hydrolases; C021480:kynureninase; D008190:Lyases; D013437:Carbon-Sulfur Lyases; C019969:S-alkylcysteine lyase