Mechanism of reactions catalyzed by selenocysteine beta-lyase.
The reaction mechanism of selenocystine beta-lyase has been studied and it was found that elemental selenium is released enzymatically from selenocysteine, and reduced to H2Se nonenzymatically with dithiothreitol or some other reductants that are added to prepare selenocysteine from selenocystine in the anaerobic reaction system. 1H and 13C NMR spectra of L-alanine formed in 2H2O have shown that an equimolar amount of [beta-2H1]- and [beta-2H2]alanines are produced. The deuterium isotope effect at the alpha position was observed; kH/kD = 2.4. These results indicated that the alpha hydrogen of selenocysteine was removed by a base at the active site, and was incorporated into the alpha position of alanine, a product, without exchange of a solvent deuterium. When the enzyme was incubated with L-selenocysteine in the absence of added pyridoxal 5'-phosphate, the activity decreased with prolonged incubation time. However, the activity was recovered by addition of 5'-phosphate. The spectrophotometric study showed that the inactivated enzyme was the apo form. The apoenzyme was activated by a combination of pyridoxamine 5'-phosphate and various alpha-keto acids such as alpha-ketoglutarate and pyruvate. Thus, the enzyme is inactivated through transamination between selenocysteine and the bound pyridoxal 5'-phosphate to produce pyridoxamine 5'-phosphate and a keto acid derived from selenocysteine. The pyridoxal enzyme, an active form, is regenerated by addition of alpha-keto acids. This regulatory mechanism is analogous to those of aspartate beta-decarboxylase [EC 4.1.1.12], arginine racemase [EC 5.1.1.9], and kynureninase [EC 3.7.1.3] [K. Soda and K. Tanizawa (1979) Adv. Enzymol. 49, 1].
Esaki
N
N
Karai
N
N
Nakamura
T
T
Tanaka
H
H
Soda
K
K
D000409:Alanine / Q000096:biosynthesis; D000818:Animals; D001665:Binding Sites; D003545:Cysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D066298:In Vitro Techniques; D007651:Keto Acids / Q000494:pharmacology; D008099:Liver / Q000201:enzymology; D008190:Lyases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011733:Pyridoxamine / Q000031:analogs & derivatives / Q000494:pharmacology; D012643:Selenium / Q000494:pharmacology; D017279:Selenocysteine; D013552:Swine
D007651:Keto Acids; D017279:Selenocysteine; D011733:Pyridoxamine; D008190:Lyases; C034278:selenocysteine lyase; D012643:Selenium; D003545:Cysteine; D000409:Alanine; C010627:pyridoxamine phosphate