Coenzyme stimulation of isomerase activity of sepiapterin reductase in the biosynthesis of tetrahydrobiopterin.
The 6-lactoyl tetrahydropterin (C1'-keto PH4) isomerase activity of sepiapterin reductase, which was found in our recent work (Katoh and Sueoka (1987) J. Biochem. 101, 275-278) as a novel activity of the enzyme, i.e., the conversion of C1'-keto PH4 to 6-1'-hydroxy-2'-oxopropyl tetrahydropterin (C2'-keto PH4) without coenzymes, could be enhanced by a small amount of NADPH or NADP+. The concentration of NADP+ required for the maximal stimulation was approximately the same as the concentration of the enzyme subunit. When NADP+ was added with the enzyme and C1'-keto PH4 at pH 8.6, the reaction sequence of C1'-keto PH4----C2'-keto PH4----tetrahydrobiopterin (BH4) was observed in the presence of dithioerythritol. These observations suggest that the coenzyme stimulating the isomerase function of sepiapterin reductase may be involved in the two sequential reductions, from pyruvoyl tetrahydropterin to BH4, by causing internal rearrangement of the keto group of the first intermediate, C1'-keto PH4, to form the second one, C2'-keto PH4.
Katoh
S
S
Department of Biochemistry, Josai Dental University, Saitama.
Sueoka
T
T
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000818:Animals; D001708:Biopterins / Q000031:analogs & derivatives* / Q000096:biosynthesis; D003067:Coenzymes / Q000378:metabolism*; D004789:Enzyme Activation; D004912:Erythrocytes / Q000201:enzymology / Q000378:metabolism; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D009249:NADP / Q000378:metabolism; D051381:Rats
D003067:Coenzymes; D001708:Biopterins; D009249:NADP; D000429:Alcohol Oxidoreductases; C022630:sepiapterin reductase; D007535:Isomerases; C003402:sapropterin