Three human alcohol dehydrogenase subunits: cDNA structure and molecular and evolutionary divergence.
Three human alcohol dehydrogenase subunits: cDNA structure and molecular and evolutionary divergence.
复制标题
三个人类乙醇脱氢酶亚基：cDNA 结构以及分子和进化分歧。
DOI：
10.1073/pnas.83.3.634
复制
发表时间：
1986
影响因子：
11.1
通讯作者：
Yoshida,A
中科院分区：
文献类型：
--
作者：
Ikuta,T;Szeto,S;Yoshida,A
I 类人乙醇脱氢酶（ADH；乙醇：NAD+ 氧化还原酶，EC 1.1.1.1）由几个由 ADH1、ADH2 和 ADH3 位点控制的 α、β 和 γ 亚基同二聚体和异二聚体组成。我们之前克隆了β亚基的全长cDNA，并建立了374个氨基酸残基的完整序列。克隆并表征了 α 和 γ 亚基的 cDNA。使用与 γ 序列匹配但不与 β 序列匹配的合成寡核苷酸探针筛选在噬菌体 lambda gt11 中构建的人肝脏 cDNA 文库。克隆pUCADH gamma 21和克隆pUCADH α 15L在限制性位点和与核苷酸探针的杂交方面与β cDNA不同。克隆pUCADH gamma 21包含1.5千碱基对(kbp)的插入并编码与报道的γ亚基的氨基酸序列相容的374个氨基酸残基。克隆pUCADH α 15L含有2.4 kbp的插入并包含编码另一个亚基（α亚基）的374个氨基酸残基的核苷酸序列。此外，该克隆包含编码 β 亚基延伸 5' 区域 COOH 末端部分的序列。三个亚基的 cDNA 的氨基酸序列和编码区非常相似（大约 93-95% 同一性）。在它们的 3' 非编码区中也观察到高度的相似性。然而，46位Zn结合半胱氨酸残基附近存在明显差异，即α中的Cys-Gly-Thr、野生型β1中的Cys-Arg-Thr、东方型β2中的Cys-His-Thr和γ中的Cys-Arg-Ser，反映了它们的动力学性质的差异。根据三个亚基的cDNA序列和推导的氨基酸序列，讨论了它们的结构和进化关系。
Class I human alcohol dehydrogenase (ADH; alcohol:NAD+ oxidoreductase, EC 1.1.1.1) consists of several homo- and heterodimers of alpha, beta, and gamma subunits that are governed by the ADH1, ADH2, and ADH3 loci. We previously cloned a full length of cDNA for the beta subunit, and the complete sequence of 374 amino acid residues was established. cDNAs for the alpha and gamma subunits were cloned and characterized. A human liver cDNA library, constructed in phage lambda gt11, was screened by using a synthetic oligonucleotide probe that was matched to the gamma but not to the beta sequence. Clone pUCADH gamma 21 and clone pUCADH alpha 15L differed from beta cDNA with respect to restriction sites and hybridization with the nucleotide probe. Clone pUCADH gamma 21 contained an insertion of 1.5 kilobase pairs (kbp) and encodes 374 amino acid residues compatible with the reported amino acid sequence of the gamma subunit. Clone pUCADH alpha 15L contained an insertion of 2.4 kbp and included nucleotide sequences that encode 374 amino acid residues for another subunit, the alpha subunit. In addition, this clone contained the sequences that encode the COOH-terminal part of the beta subunit at its extended 5' region. The amino acid sequences and coding regions of the cDNAs of the three subunits are very similar (approximately 93-95% identity). A high degree of resemblance is observed also in their 3' noncoding regions. However, distinctive differences exist in the vicinity of the Zn-binding cysteine residue at position 46--i.e., Cys-Gly-Thr in the alpha, Cys-Arg-Thr in the wild-type beta 1, Cys-His-Thr in the Oriental-type beta 2, and Cys-Arg-Ser in the gamma, reflecting the differences in their kinetic properties. Based on the cDNA sequences and the deduced amino acid sequences of the three subunits, their structural and evolutionary relationships are discussed.