Human peptidylarginine deiminase type II: molecular cloning, gene organization, and expression in human skin
Human peptidylarginine deiminase type II: molecular cloning, gene organization, and expression in human skin
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DOI：
10.1016/s0003-9861(02)00516-7
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发表时间：
2002-11-01
影响因子：
3.9
通讯作者：
Maruyama, N
中科院分区：
文献类型：
--
作者：
Ishigami, A;Ohsawa, T;Maruyama, N
肽基精氨酸脱亚胺酶（PADS）是翻译后修饰酶，以钙离子依赖性方式将蛋白质精氨酸转化为瓜氨酸残基。啮齿动物有四种PAD亚型（I、II、III和IV型），每种亚型的底物和组织特异性不同。事实上，唯一检测到所有四种PAD mRNA的组织是表皮。在这项研究中，我们发现在HSC-1人皮肤鳞状细胞癌细胞在体外的PAD活性，这种活性在培养过程中增加。使用同源性为基础的策略，我们克隆了全长cDNA编码人PAD II型。cDNA长2348，编码665个氨基酸的序列，预测分子量为75 kDa。预测的蛋白质与大鼠和小鼠PAD II型序列具有93%的同一性。从这两个物种的氨基酸序列的比对显示显着的保守在C-末端区域，这表明存在的功能区域，如酶催化位点和/或钙结合域。基因组学分析表明，人PAD II型位于染色体1p35.2-p35.21，全长50 kb，包含16个外显子和15个内含子。随后在大肠杆菌中产生的重组PAD蛋白被证明是酶促活性的，具有与大鼠PAD II型相似的底物特异性。在人类皮肤的免疫组织化学研究中，II型酶表达的所有活的表皮层，表明PAD 11型是表皮角质形成细胞的终末分化过程中的重要功能。(C)2002 Elsevier Science（美国）。All rights reserved.
Peptidylarginine deiminases (PADS) are posttranslational modification enzymes that convert protein arginine to citrulline residues in a calcium ion-dependent manner. Rodents have four isoforms of PAD (types I, II, III, and IV), each of which is distinct in substrate and tissue specificity. In fact, the only tissue in which all four PAD mRNAs have been detected is the epidermis. In this study, we found PAD activity in HSC-1 human cutaneous squamous carcinoma cells in vitro, and this activity increased during cultivation. Using a homology-based strategy, we cloned a full-length cDNA encoding human PAD type II. The cDNA was 2348 by long and encoded a 665-amino-acid sequence with a predicted molecular mass of 75 kDa. The predicted protein shared 93% identity with the rat and mouse PAD type II sequence. Alignment of the amino acid sequences from both species revealed notable conservation in the C-terminal region, suggesting the presence of a functional region such as an enzyme catalytic site and/or a calcium-binding domain. Gene organization analysis established that human PAD type II on chromosome 1p35.2-p35.21 spanned more than 50 kb and contained 16 exons and 15 introns. A recombinant PAD protein subsequently produced in Escherichia coli proved to be enzymatically active, with substrate specificities similar to those of the rat PAD type II. In an immunohistochemical study of human skin, the type II enzyme was expressed by all the living epidermal layers, suggesting that PAD type 11 is functionally important during terminal differentiation of epidermal keratinocytes. (C) 2002 Elsevier Science (USA). All rights reserved.