Davis M D, Ribeiro P, Tipper J, Kaufman S
Laboratory of Neurochemistry, National Institute of Mental Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10109-13. doi: 10.1073/pnas.89.21.10109.
The ability of 2-amino-4-hydroxy-7-[dihydroxylpropyl-(L-erythro)-5,6,7,8-tetrahyd ropterin] ("7-tetrahydrobiopterin" or 7-BH4) to substitute for the natural cofactor tetrahydrobiopterin (BH4) has been studied in vitro in the reactions of the three mammalian aromatic amino acid hydroxylases. With rat liver phenylalanine hydroxylase, the apparent Km for 7-BH4 is 160 microM, a value that is approximately 60-fold greater than that for the natural cofactor. In contrast, the hydroxylase reaction is severely inhibited by as little as 1 microM 7-BH4 when assayed in the presence of physiological concentrations of BH4. This inhibition can be overcome either by an increase in the concentration of BH4 or a decrease in the concentration of phenylalanine. With both rat brain tryptophan hydroxylase and rat pheochromocytoma tyrosine hydroxylase, the Km value for 7-BH4 is about one order of magnitude greater than the Km for BH4. Accordingly, 7-BH4 is a poor competitive inhibitor of both tryptophan and tyrosine hydroxylase. Thus, our results suggest that the observed hyperphenylalaninemia in patients who excrete 7-BH4 in their urine may arise directly from the inhibition of phenylalanine hydroxylase by low levels of this pterin. On the other hand, it is less likely that low levels of 7-BH4 would affect the activity of tyrosine or tryptophan hydroxylase in vivo.
在三种哺乳动物芳香族氨基酸羟化酶的反应中,已在体外研究了2-氨基-4-羟基-7-[二羟基丙基-(L-赤藓糖型)-5,6,7,8-四氢蝶呤](“7-四氢生物蝶呤”或7-BH4)替代天然辅因子四氢生物蝶呤(BH4)的能力。对于大鼠肝脏苯丙氨酸羟化酶,7-BH4的表观Km为160μM,该值约为天然辅因子的60倍。相反,当在生理浓度的BH4存在下进行测定时,低至1μM的7-BH4就能严重抑制羟化酶反应。这种抑制作用可通过增加BH4的浓度或降低苯丙氨酸的浓度来克服。对于大鼠脑色氨酸羟化酶和大鼠嗜铬细胞瘤酪氨酸羟化酶,7-BH4的Km值比BH4的Km值大约高一个数量级。因此,7-BH4是色氨酸和酪氨酸羟化酶的弱竞争性抑制剂。所以,我们的结果表明,尿中排泄7-BH4的患者中观察到的高苯丙氨酸血症可能直接源于这种蝶呤水平较低对苯丙氨酸羟化酶的抑制。另一方面,低水平的7-BH4在体内影响酪氨酸或色氨酸羟化酶活性的可能性较小。