Studies on the structure and biological function of human hair follicle peptidylarginine deiminase

毛发由毛母细胞分化而来的内、根鞘、角质层和髓质细胞组成。毛母细胞的终末分化是示踪表皮基底细胞的常见方式，但有关毛发分化过程的生化研究很少。多肽精氨酸脱亚胺酶(PAD)是一种翻译后修饰酶，通过将目标蛋白的碱性区域转化为放射性价态来产生静电变化，导致目标蛋白与另一种蛋白之间的不同相互作用。自从1977年G.E.Rogers首次发现酶活性将豚鼠毛囊提取液中蛋白质的精氨酸残基转化为瓜氨酸残基以来，PAD的生化研究在各种组织中都取得了进展。然而，对毛囊垫的研究却很少。在本研究中，我们采用原代培养的方法制备了人毛细胞，并进行了rna，然后克隆了enti基因的…。通过同源聚合酶链式反应获得更多的人毛细胞PAD(Pad T4)的Re基因。通过核苷酸序列分析，确定了完整的氨基酸序列。我们在大肠杆菌中构建了人Pad T4的表达载体，并对重组酶的纯化和酶学性质进行了研究。特别是，毛细胞特有的蛋白(Trichohyalin)对毛发结构的调节具有重要作用，并且在Trichohyalin中发现了瓜氨酸残基，因此Trichohyalin是PAD T4的候选靶标。在本研究中，我们证明了Pad T4有效地修饰了曲霉素A。接下来，我们制备了针对Pad T4 IS的单抗，并检测了Pad T4在毛囊中的细胞分布。免疫组织化学染色显示PAD T4定位于毛细胞内根鞘。证明上述赤霉素酶在局部也存在于该组织中的知识，以及该酶使曲霉素林成为靶子的知识。较少

The hair is composed of inner and root sheath, cuticle, and medulla cells which are differentiated from hair mother cell. The terminal differentiation of hair mother cell is common in tracing epidermal basal cells, but the biochemical study regarding the process of hair differentiation is very few.Peptidylarginine deiminase(PAD) is a post-translational modification enzyme which gives the electrostatic change by converting basic region of the target protein into the radio valence, causing the different interaction between target proteins and another protein. Since the first finding of G.E Rogers in 1977, that enzymatic activity converts arginine residue of the protein to the citrulline residue in extract of guinea pig hair follicle, biochemical studies of PADs were advanced in the various tissues. However the research on the PAD of the hair follicle is seldom carried out. In this study, we prepared the human hair cells by the primary culture method, and the RNA, and then cloned the enti … More re cDNA of the human hairy cell PAD(PAD T4) by homologue polymerase chain reaction. The complete amino acid sequence was clarified by the nucleotide sequence analysis. We constructed the human PAD T4 expression plasmid in an Echerichia coli and purification and enzymatic properties of the recombinant enzyme were clarified. Especially, the role of which the hairy cell peculiar protein(trichohyalin) is important for the regulation of the hair construct and the citrulline residues were found in the trichohyalin, that the trichohyalin is a candidate target for PAD T4. In this study we proved that PAD T4 modified the trichohyalin effectively. Next, we made a monoclonal antibody against specific for PAD T4 is and we examined the cellular distribution of PAD T4 in the hair follicle. The immunohistochemisty showed that the PAD T4 specifically localized in the inner root sheath in the hairy cells. The knowledge which proved that the above-mentioned trichohyalin also exists in this organization locally and that this enzyme has made the trichohyalin to be a target. Less