Isolation of cDNA for human epidermal type I transglutaminase.
Isolation of cDNA for human epidermal type I transglutaminase.
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人表皮 I 型转谷氨酰胺酶 cDNA 的分离。
DOI：
10.1111/1523-1747.ep12464554
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Goldsmith,LA
中科院分区：
文献类型：
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作者：
Polakowska,R;Herting,E;Goldsmith,LA
复层上皮（包括表皮）的角质形成细胞表达两种不同形式的转氨酶，I型和II型。I型转氨酶活性负责终末分化培养的角质形成细胞中的细胞包膜形成。转氨酶的酶活性与体内和体外差异表达的几种蛋白质有关。为了阐明表皮表达的转氨酶之间的关系，从人高钙角质形成细胞λ gt 11文库中克隆了I型转氨酶的cDNA。使用由PCR引发的cDNA片段作为筛选探针，所述PCR引发用编码活性位点中的五个不变氨基酸的寡核苷酸的混合物。基于λ1- 126 a克隆中1653 nt的序列分析和互补的约3 kb转录本的表达模式，我们报告了表皮I型转氨酶基因的克隆。在培养的人角质形成细胞中，该基因的表达受钙离子和视黄酸的调节。在活性位点半胱氨酸残基附近有高度保守的区域，这可能对酶的特殊功能很重要。
Keratinocytes of stratified epithelia, including the epidermis, express two distinct forms of transglutaminase, type I and type II. Type I transglutaminase activity is responsible for cell envelope formation in terminally differentiating cultured keratinocytes. Transglutaminase enzymatic activity has been associated with several proteins that are differentially expressed in vivo and in vitro. To elucidate the relationship between the epidermally expressed transglutaminases, cDNA for type I transglutaminase was cloned from a human high-calcium keratinocyte λgt11 library. cDNA fragments, generated by PCR primed with a mixture of oligonucleotides coding for five invariant amino acids in the active site, were used as a screening probe. Based on the sequence analysis of 1653 nt contained in the λ1–126a clone and on the pattern of expression of a complementary approximately 3-kb transcript, we report cloning of the epidermal type I transglutaminase gene. The expression of this gene is regulated by calcium ions and retinoic acid in cultured human keratinocytes. There are highly conserved regions near the active site cysteine residues that may be important for the enzyme's specialized functions.