大鼠肝脏蝶呤还原酶cDNA的分离与表达
Isolation and expression of rat liver sepiapterin reductase cDNA.
作者信息
Citron B A, Milstien S, Gutierrez J C, Levine R A, Yanak B L, Kaufman S
机构信息
Laboratory of Neurochemistry, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892.
出版信息
Proc Natl Acad Sci U S A. 1990 Aug;87(16):6436-40. doi: 10.1073/pnas.87.16.6436.
Sepiapterin reductase (7,8-dihydrobiopterin: NADP+ oxidoreductase, EC 1.1.1.153) catalyzes the terminal step in the biosynthetic pathway for tetrahydrobiopterin, the cofactor necessary for aromatic amino acid hydroxylation. We report here the isolation of a cDNA clone for rat liver sepiapterin reductase. The cDNA has been excised from a lambda vector and the DNA sequence was determined. The insert contains the coding sequence for at least 95% of the rat enzyme and is fused to the Escherichia coli beta-galactosidase N-terminal segment and the lac promoter. The N-terminal region of the clone contains an extraordinarily high G + C content. The amino acid sequence deduced from the clone is in agreement with the size and composition of the enzyme and was matched to several tryptic peptide sequences. The enzyme encoded by the cDNA insert was shown to have sepiapterin reductase activity after expression in E. coli. Structural similarities were identified between this protein and several enzymes that should contain similar nucleotide and pteridine binding sites.
蝶呤还原酶（7,8-二氢生物蝶呤：NADP +氧化还原酶，EC 1.1.1.153）催化四氢生物蝶呤生物合成途径的最后一步，四氢生物蝶呤是芳香族氨基酸羟化所必需的辅因子。我们在此报告大鼠肝脏蝶呤还原酶cDNA克隆的分离。该cDNA已从λ载体中切出并测定了DNA序列。插入片段包含大鼠酶至少95%的编码序列，并与大肠杆菌β-半乳糖苷酶N端片段和lac启动子融合。该克隆的N端区域含有异常高的G + C含量。从该克隆推导的氨基酸序列与该酶的大小和组成一致，并与几个胰蛋白酶肽序列相匹配。在大肠杆菌中表达后，由cDNA插入片段编码的酶显示具有蝶呤还原酶活性。在该蛋白质与几种应含有相似核苷酸和蝶呤结合位点的酶之间鉴定出结构相似性。
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