Ceramides are bound to structural proteins of the human foreskin epidermal cornified cell envelope
Ceramides are bound to structural proteins of the human foreskin epidermal cornified cell envelope
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DOI：
10.1074/jbc.273.28.17763
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发表时间：
1998-07-10
影响因子：
4.8
通讯作者：
Steinert, PM
中科院分区：
文献类型：
--
作者：
Marekov, LN;Steinert, PM
人类表皮屏障功能的一个重要组成部分是神经酰胺，神经酰胺通过酯键与角化细胞膜(CE)的未知蛋白质结合，在本文中，我们研究了神经酰胺附着的蛋白质靶标。通过对分离的包皮表皮细胞进行部分皂化，然后进行有限的蛋白降解，我们回收了几种脂肽。生化和质谱学表征表明，它们都含有接近化学计量质量的神经酰胺，质量从690到890个原子质量单位不等，其中常见的六种主要定量物种。神经酰胺序列与已知的猪皮神经酰胺的组成相似，从而为其脂肪酸和鞘氨醇的组成提供了强有力的间接数据，回收的多肽约占包皮CE神经酰胺总量的20%，通过氨基酸序列分析，约35%的多肽来自于重要的包皮CE结构蛋白--总蛋白的祖先谷氨酰胺富集区。另有18%来源于周斑蛋白和包膜蛋白的杆状结构域序列，这两种蛋白也是已知或可疑的CE蛋白。其他的多肽太短，难以明确识别。综上所述，这些数据表明，包衣蛋白、包膜蛋白、包膜蛋白以及可能的其他结构蛋白作为底物，通过酯键将神经酰胺连接到CE上，以实现人体表皮的屏障功能。
An important component of barrier function in human epidermis is contributed by ceramides that are bound by ester linkages to undefined proteins of the cornified cell envelope (CE), In this paper, we have examined the protein targets for the ceramide attachment. By partial saponification of isolated foreskin epidermal CEs followed by limited proteolysis, we have recovered several lipopeptides. Biochemical and mass spectroscopic characterization revealed that all contained near stoichiometric amounts of ceramides of masses ranging from about 690 to 890 atomic mass units, of which six quantitatively major species were common. The array of ceramides was similar to that obtained from pig skin, the composition of which is known, thereby providing strong indirect data for their fatty acid and sphingosine compositions, The recovered peptides accounted for about 20% of the total foreskin CE ceramides, By amino acid sequencing, about 35% of the peptides were derived from ancestral glutamine glutamate-rich regions of involucrin, an important CE structural protein. Another 18% derived from rod domain sequences of periplakin and envoplakin, which are also known or suspected CE proteins. Other peptides were too short for unequivocal identification. Together, these data indicate that involucrin, envoplakin, periplakin, and possibly other structural proteins serve as substrates for the attachment of ceramides by ester linkages to the CE for barrier function in human epidermis.