CHARACTERIZATION OF SCIELLIN, A PRECURSOR TO THE CORNIFIED ENVELOPE OF HUMAN KERATINOCYTES
CHARACTERIZATION OF SCIELLIN, A PRECURSOR TO THE CORNIFIED ENVELOPE OF HUMAN KERATINOCYTES
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DOI：
10.1111/j.1432-0436.1992.tb00667.x
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发表时间：
1992-04-01
期刊：
影响因子：
2.9
通讯作者：
BADEN, HP
中科院分区：
文献类型：
--
作者：
KVEDAR, JC;MANABE, M;BADEN, HP
角化包膜位于终末分化的角质形成细胞的质膜下，是由膜相关的谷氨酰胺转胺酶交联的蛋白质前体形成的。这份报告描述了一种新的凝固信封的前兆。用人培养的角质形成细胞的角质化包膜免疫小鼠获得的单克隆抗体可染色表皮和其他层状鳞状上皮（包括头发和指甲）中分化程度较高的细胞的周围。免疫组化和Western分析表明，该抗原表位在哺乳动物中具有广泛的保守性。免疫电镜将表位定位在表皮棘层和颗粒层的细胞外周。在毛囊中，表位存在于内根鞘和外根鞘最里面的胚芽中。该抗体识别出一个相对迁移率（M(r)） 82000, pl 7.8的蛋白。该蛋白是转谷氨酰胺酶的底物，如丹西尸碱掺入试验所示。纯化的角质包膜吸收了抗体对部分纯化蛋白的反应性，溴化氰切割包膜导致免疫反应片段的释放。该蛋白仅溶于8 M尿素或2%十二烷基硫酸钠（SDS）等变性缓冲液中。在50 mM TRIS pH 8.3 + 0.3 M NaCl（高盐缓冲液）中可实现部分溶解度；还原剂的存在不影响溶解度。在8 M尿素中提取培养的角质形成细胞，随后在50 mM TRIS pH 8.3缓冲液中透析，导致蛋白与角蛋白丝沉淀。高盐缓冲液的透析阻止了蛋白质的沉淀。这种蛋白质独特的溶解度表明，它与自身和/或与角蛋白丝聚集。讨论了该蛋白在凝固包膜组装中的可能作用。我们将这种蛋白质命名为Sciellin（来自古英语“sciell”，意为壳）。
The cornified envelope, located beneath the plasma membrane of terminally differentiated keratinocytes, is formed as protein precursors are cross-linked by a membrane associated transglutaminase. This report characterizes a new precursor to the cornified envelope. A monoclonal antibody derived from mice immunized with cornified envelopes of human cultured keratinocytes stained the periphery of more differentiated cells in epidermis and other stratified squamous epithelia including hair and nails. The epitope was widely conserved among mammals as determined by immunohistochemical and Western analysis. Immunoelectron microscopy localized the epitope to the cell periphery in the upper stratum spinosum and granulosum of epidermis. In the hair follicle, the epitope was present in the internal root sheath and in the infundibulum, the innermost aspect of the external root sheath. The antibody recognized a protein of relative mobility (M(r)) 82,000, pl 7.8. The protein was a transglutaminase substrate as shown by a dansylcadaverine incorporation assay. Purified cornified envelopes absorbed the reactivity of the antibody to the partially purified protein and cleavage of envelopes by cyanogen bromide resulted in release of immunoreactive fragments. The protein was soluble only in denaturing buffers such as 8 M urea or 2% sodium dodecyl-sulfate (SDS). Partial solubility could be achieved in 50 mM TRIS pH 8.3 plus 0.3 M NaCl (high salt buffer); the presence of a reducing agent did not affect solubility. Extraction of cultured keratinocytes in 8 M urea and subsequent dialysis against 50 mM TRIS pH 8.3 buffer resulted in precipitation of the protein with the keratin filaments. Dialysis against high salt buffer prevented precipitation of the protein. The unique solubility properties of this protein suggest that it aggregates with itself and/or with keratin filaments. The possible role of the protein in cornified envelope assembly is discussed. We have named this protein Sciellin (from the old english "sciell" for shell).