Robson K J, Beattie W, James R J, Cotton R C, Morgan F J, Woo S L
Biochemistry. 1984 Nov 20;23(24):5671-5. doi: 10.1021/bi00319a001.
Classical phenylketonuria, an inborn error in metabolism, is caused by a deficiency of the hepatic enzyme phenylalanine hydroxylase. The identification of putative cDNA clones coding for rat liver phenylalanine hydroxylase by hybrid-selected translation has previously been reported [Robson, K. J., Chandra, T., MacGillivray, R. T. A., & Woo, S. L. C. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 4701-4705]. The authenticity of the clones, however, could not be definitively ascertained at the time because of a lack of amino acid sequence data of the enzyme in the literature. Purified rat liver phenylalanine hydroxylase was subjected to cyanogen bromide treatment, and the resulting fragments were used for N-terminal amino acid sequence analysis. The partial amino acid sequence was then compared to that deduced from an open reading frame in the nucleotide sequence of the cDNA clones. A perfect match of 17 amino acid residues was found between the two sequences following a unique methionine codon present in the nucleotide sequence, thereby providing unambiguous evidence for the identity of the rat liver phenylalanine hydroxylase cDNA clones.
经典型苯丙酮尿症是一种先天性代谢缺陷疾病，由肝脏酶苯丙氨酸羟化酶缺乏引起。此前已有报道通过杂交选择翻译鉴定出编码大鼠肝脏苯丙氨酸羟化酶的假定cDNA克隆[罗布森，K. J.，钱德拉，T.，麦吉利夫雷，R. T. A.，& 吴，S. L. C.（1982年）《美国国家科学院院刊》79，4701 - 4705]。然而，由于当时文献中缺乏该酶的氨基酸序列数据，这些克隆的真实性无法得到确切证实。对纯化的大鼠肝脏苯丙氨酸羟化酶进行溴化氰处理，所得片段用于N端氨基酸序列分析。然后将部分氨基酸序列与从cDNA克隆核苷酸序列中的开放阅读框推导出来的序列进行比较。在核苷酸序列中存在的一个独特甲硫氨酸密码子之后，发现两个序列之间有17个氨基酸残基完全匹配，从而为大鼠肝脏苯丙氨酸羟化酶cDNA克隆的同一性提供了明确证据。
