Mizutani M, Ward E, DiMaio J, Ohta D, Ryals J, Sato R
International Research Laboratories, Ciba-Geigy Japan Ltd., Takarazuka, Japan.
Biochem Biophys Res Commun. 1993 Feb 15;190(3):875-80. doi: 10.1006/bbrc.1993.1130.
With the aid of partial amino acid sequences determined for cinnamate 4-hydroxylase (P450C4H) purified from mung bean seedlings, two cDNA clones were isolated and their inserts were completely sequenced. The nucleotide sequences of the two clones were nearly identical and contained an open reading frame predicted to encode a polypeptide consisting of 505 amino acid residues. The partial sequences determined from the purified P450C4H closely corresponded to the primary structures deduced from the cDNA sequences. This is the first isolation of cDNA clones encoding a higher plant P450 possessing clear physiological activity. Comparison to known cytochromes P450 indicated that P450C4H belongs to a novel P450 gene family.
借助从绿豆幼苗中纯化得到的肉桂酸4-羟化酶(P450C4H)所测定的部分氨基酸序列,分离出了两个cDNA克隆,并对其插入片段进行了全序列测定。这两个克隆的核苷酸序列几乎完全相同,包含一个开放阅读框,预计可编码一个由505个氨基酸残基组成的多肽。从纯化的P450C4H中测定的部分序列与从cDNA序列推导的一级结构紧密对应。这是首次分离出编码具有明确生理活性的高等植物P450的cDNA克隆。与已知的细胞色素P450比较表明,P450C4H属于一个新的P450基因家族。