Aoki N, Ito K, Ito M
Department of Dermatology, Niigata University School of Medicine, Japan.
J Invest Dermatol. 1998 Nov;111(5):804-9. doi: 10.1046/j.1523-1747.1998.00387.x.
To identify the anagen hair follicle-specific proteins, we screened the cDNA library prepared from the murine skins of anagen phase by the differential hybridization technique. Fifty-four cDNA clones expressed specifically in anagen phase were isolated, and most were found to correspond to known proteins in the hair follicles. Alternatively, we isolated a cDNA clone encoding a novel protein that possessed an entire open reading frame of 501 base pairs. This protein with a molecular weight of 17.9 kDa has no specific motifs nor significant homology to proteins already reported, although it contains some direct repeats that are often observed in intermediate filament-associated proteins and has a similar amino acid composition as a member of them. Northern blot analysis demonstrates that the transcript of this protein is skin specific, and that it is present in mid- and late anagen but not in catagen, telogen, and early anagen phases. The transcript appears to be expressed specifically in the keratogenous zone of the cortical cells of hair follicles, as exhibited by in situ hybridization. Furthermore, immunohistochemical study confirms that the protein is distributed in the cytoplasms of the keratinizing cortical cells and is undetected in the completely keratinized ones. These results suggest that this protein can be identified as a new member of intermediate filament-associated proteins and is related to the keratinization of the cortical cell layer in mouse hair. Therefore, we have termed this novel protein mKAP13 according to the unified nomenclature.
为了鉴定生长期毛囊特异性蛋白,我们采用差异杂交技术筛选了从生长期小鼠皮肤制备的cDNA文库。分离出54个在生长期特异性表达的cDNA克隆,发现大多数与毛囊中的已知蛋白相对应。另外,我们分离出一个编码新蛋白的cDNA克隆,该蛋白具有501个碱基对的完整开放阅读框。这种分子量为17.9 kDa的蛋白没有特定的基序,与已报道的蛋白也没有显著同源性,尽管它包含一些在中间丝相关蛋白中经常观察到的直接重复序列,并且氨基酸组成与其中一个成员相似。Northern印迹分析表明,该蛋白的转录本是皮肤特异性的,并且存在于生长期中期和后期,而在退行期、休止期和生长期早期不存在。原位杂交显示,该转录本似乎在毛囊皮质细胞的角质形成区特异性表达。此外,免疫组织化学研究证实,该蛋白分布在角质化皮质细胞的细胞质中,在完全角质化的细胞中未检测到。这些结果表明,该蛋白可被鉴定为中间丝相关蛋白的一个新成员,并且与小鼠毛发皮质细胞层的角质化有关。因此,我们根据统一命名法将这种新蛋白命名为mKAP13。