New insights into the active center of rat liver cystathionase.
Treatment by urea of purified rat liver cystathionase (L-Cystathionine cysteine-lyase (deaminating), EC 4.4.1.1) provoked a similar alteration of two activities of the enzyme, namely cysteine desulfhydration and homoserine deamination. Since the decreases of the two activities were also comparable as a result of chymotrypsin digestion of the enzyme, these observations suggest that the two sites responsible for the one and the other activites are in close proximity. Studies of the effect of derivatives of substrates (S-carboxymethylcyste-ine, S-carboxyethylcysteine, S-carboxymethylhomocysteine and S-carboxyethylhomocysteine) on both activities were performed. All of them inhibited cysteine desulfhydration and homoserine deamination; in several cases, the type of inhibition was also determined. The results are in agreement with the hypothesis that each of the two sites of the active center has, at least, three binding points which "recognise" groupings of substrates or of inhibitors, and this led us to propose a model for the active center. Each site has an -NH-2 binding point, hence the active center has two -NH-2 binding points; therefore, as cystathionase consists of four subunits and contains four molecules of pyriodoxal phosphate, it might be of interest to determine whether the smallest active molecule is the dimer.
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002918:Chymotrypsin / Q000494:pharmacology; D003540:Cystathionine / Q000031:analogs & derivatives; D003542:Cystathionine gamma-Lyase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D006714:Homoserine / Q000031:analogs & derivatives / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008190:Lyases / Q000378:metabolism*; D011487:Protein Conformation; D051381:Rats; D013329:Structure-Activity Relationship; D014508:Urea / Q000494:pharmacology
D003540:Cystathionine; D006714:Homoserine; D014508:Urea; D002918:Chymotrypsin; D008190:Lyases; D003542:Cystathionine gamma-Lyase