Relationship of atypical phenylketonuria and a new type of carbonyl reductase which related to synthesize BH4

非典型苯丙酮尿症与合成BH4的新型羰基还原酶的关系

• 批准号: 12670785
• 负责人: IINO Teruhiko
• 金额: $ 2.3万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670785/
• 关键词: Tetrahydrobiopterin; Carbonyl reductase; Biosynthesis of BH4; BH4 deficiency; BH_4生合成系; BH_4欠損症; セピアプテリン還元酵素; カイコ; 異型フェニルケトン尿症

Tetrahydrobiopterin plays an important role in the biosynthesis of certain neurotransmitters. Using DEAE-Sepharose FF column chromatography, we separated the enzyme that synthesizes tetrahydrobiopterin(BH4) from 6- pyruvoyl-tetrahydropterin(PPH4), which is different from sepiapterin reductase, in the lemon mutant of the silkworm, Bombyx mori , into two fractions, which were named carbonyl reductase I ( CR I ) and carbonyl reductase II ( CR II ). When CR I was mixed with CR II in the reaction mixture, PPH4 was reduced to BH4 in the presence of NADPH.Furthermore, we examined the formation of BH4 from PPH4 by several human carbonyl reductases which belong to the aldo-keto reductase ( AKR1) family members. In these experiments, AKR1A1, AKR1B1, AKR1C1, AKR1C2, and AKR1C4 converted PPH4 to 1'-oxo-2'-hydroxypropyl-tetrahydropterin, while AKR1C3 converted PPH4 to 1'-hydroxy-2'-oxopropyl-tetrahydropterin, both reactions occurred only in the presence of NADPH. Neither of these carbonyl reductases alone was able to catalyze the conversion of PPH4 to BH4 in the presence of NADPH.

四氢生物蝶呤在某些神经递质的生物合成中起重要作用。采用DEAE-Sepharose FF柱层析法，将家蚕柠檬突变体中与Sepiapterin还原酶不同的6-乙酰基-四氢蝶呤（6-acetyl-tetrahydropterin，PPH_4）合成四氢生物蝶呤（tetrahydrobiopterin，BH_4）的酶分离成两个组分，分别命名为羰基还原酶Ⅰ（carbonyl reductase I，CR Ⅰ）和羰基还原酶Ⅱ（carbonyl reductase II，CR Ⅱ）。当CR Ⅰ与CR Ⅱ混合时，在NADPH存在下，PPH 4被还原为BH 4.此外，我们还研究了几种属于醛酮还原酶（AKR 1）家族成员的人羰基还原酶对PPH 4生成BH 4的作用.在这些实验中，AKR 1A 1、AKR 1B 1、AKR 1C 1、AKR 1C 2和AKR 1C 4将PPH 4转化为1 '-氧代-2'-羟丙基-四氢蝶呤，而AKR 1C 3将PPH 4转化为1 '-羟基-2'-氧代丙基-四氢蝶呤，两种反应均仅在NADPH存在下发生。这些羰基还原酶单独都不能催化NADPH存在下PPH 4转化为BH 4。

项目成果

Teruhiko Iino: "The enzyme that synthesizes tetrahydrobiopterin from 6-pyruvoyl-tetrahydropterin in the lemon mutant silkworm consists of two carbonyl reductases"Archiv. Biochem. Biophys.. 373. 442-446 (2000)

Teruhiko Iino：“柠檬突变蚕中从 6-丙酮酰四氢蝶呤合成四氢生物蝶呤的酶由两个羰基还原酶组成”Archiv。

Hiroshi Sawada: "Molecular cloning and characterization of a encoding a novel cuticle protein in the silkworm, Bombyx mori"Comp. Biochem. Biophys. Part B. (in press). (2003)

Hiroshi Sawada：“蚕 Bombyx mori 中编码新型角质层蛋白的分子克隆和表征”Comp。

Teruhiko Iino: "The Enzyme That Synthesizes Tetrahydrobiopterin from 6-pyruvoyl-tetrahydropterin in the lemon mutant Silkworm Consists of Two Carbonyl Reductases"Archives of Biochemistry and Biophysics. 373. 442-446 (2000)

Teruhiko Iino：“柠檬突变体蚕中从 6-丙酮酰四氢蝶呤合成四氢生物蝶呤的酶由两个羰基还原酶组成”生物化学和生物物理学档案。

TERUHIKO IINO: "The Enzyme that synthesizes Tetrahydrobiopterin from 6-pyruvoyl-tetrahydropterin in the lemon mutant silkworm consists of two carbonyl reductases"Arch.Biochem.Biophys.. 373. 442-446 (2000)

TERUHIKO IINO：“柠檬突变蚕中从 6-丙酮酰-四氢蝶呤合成四氢生物蝶呤的酶由两个羰基还原酶组成”Arch.Biochem.Biophys.. 373. 442-446 (2000)