cDNA cloning and characterization of sciellin, a LIM domain protein of the keratinocyte cornified envelope
cDNA cloning and characterization of sciellin, a LIM domain protein of the keratinocyte cornified envelope
复制标题
DOI：
10.1074/jbc.273.47.31547
复制
发表时间：
1998-11-20
影响因子：
4.8
通讯作者：
Olson, PF
中科院分区：
文献类型：
--
作者：
Champliaud, MF;Burgeson, RE;Olson, PF
胶原蛋白是哺乳动物角化组织角化包膜的前身。我们用一种分泌素特异性的单抗筛选人角质形成细胞表达文库，从而克隆了分泌素的编码基因。2.35kb的复合cDNA编码75.3 kDa的蛋白质，等电点为10.09，翻译的序列含有16个重复，每个重复20个氨基酸，富含Gin和Lys残基，这是潜在的转谷氨酰胺酶底物，以及一个含有单一LIM基序的羧基结构域。用荧光原位杂交技术将该基因定位于人染色体13q22带，放射杂交定位表明该基因与序列标记的位点标记Wi-457连锁，优势比对数为7.77，人包皮组织切片原位杂交证实该基因在颗粒层中表达，免疫荧光染色显示该基因定位于上层细胞。在表皮和复层扁平上皮中，如口腔、食道和阴道。除羊膜外，单层和柱状上皮未见反应。
Sciellin is a precursor of the cornified envelopes of mammalian keratinizing tissues. We have cloned the cDNA encoding sciellin by screening a human keratinocyte expression library with a sciellin-specific monoclonal antibody. The composite cDNA of 2.35 kilobase pairs encodes a protein of 75.3 kDa with a pi of 10.09, The translated sequence has a central domain containing 16 repeats of 20 amino acids each that is rich in Gin and Lys residues, which are potential transglutaminase substrates, and a carboxyl domain, which contains a single LIM motif, Sciellin cDNA probes hybridize to bands of 3.4 and 4.4 kilobase pairs on Northern blots of cultured human keratinocyte RNA. The gene was mapped to human chromosome band 13q22 by fluorescence in situ hybridization, Radiation hybrid mapping demonstrated that sciellin is linked to the sequence tagged site marker WI-457 with a logarithm of the odds score of 7.77, In situ hybridization of human foreskin tissue sections demonstrated that sciellin is expressed in the stratum granulosum, Immunofluorescent staining with a polyclonal rabbit antibody made to a recombinant sciellin protein showed peripheral cytoplasmic localization in the upper cell. layers of epidermis and in stratified squamous epithelia such as the oral cavity, esophagus, and vagina. Simple and columnar epithelia, with the exception of the amnion, showed no reaction.