Comparative proteomic profiling of murine skin
Comparative proteomic profiling of murine skin
复制标题
DOI：
10.1046/j.1523-1747.2003.12327.x
复制
发表时间：
2003-07-01
影响因子：
6.5
通讯作者：
Tang, DCC
中科院分区：
文献类型：
--
作者：
Huang, CM;Foster, KW;Tang, DCC
哺乳动物的皮肤经常暴露在不同的环境压力下，每种环境压力都会导致蛋白质表达的特定代偿性变化，这些变化可以通过蛋白质组学分析来评估。我们已经建立了BALB/c小鼠皮肤的参考蛋白质组图，允许在单个二维聚丙烯酰胺凝胶中分辨率超过500个蛋白质点。采用基质辅助激光解吸/电离飞行时间质谱法和Mascot在线数据库搜索算法，鉴定了28种不同皮肤蛋白的44个蛋白点。25种蛋白在表皮中有较高水平的表达，而只有9种蛋白主要在表皮下组织中表达。一部分蛋白斑点表现出菌株特异性表达。鉴定出多种功能的蛋白质，包括参与应激反应、细胞凋亡、生长抑制、结构完整性维持、翻译控制、能量代谢、钙结合、胆固醇运输和自由基清除的蛋白质。在皮肤中检测到Prohibitin的表达，在表皮中有更丰富的蛋白和mRNA水平。还鉴定了蛋白二硫异构酶、78 kDa葡萄糖调节蛋白前体、热休克蛋白60 （HSP60）、HSP70和HSP27等5种分子伴侣。其中，HSP27的表达主要局限于表皮，蛋白二硫异构酶的表达主要存在于表皮下组织。热休克或冷休克后皮肤的蛋白质组学分析导致HSP27、HSP60和HSP70水平升高，这表明这些伴侣蛋白参与了皮肤对温度应激的反应机制。这些数据在小鼠皮肤蛋白质组图中建立了许多参考标记，并为未来研究表皮和皮下对环境变化的反应提供了重要框架。
Mammalian skin is regularly exposed to different environmental stresses, each of which results in specific compensatory changes in protein expression that can be assessed by proteomic analysis. We have established a reference proteome map of BALB/c murine skin allowing the resolution of greater than 500 protein spots in a single two-dimensional polyacrylamide gel. Forty-four protein spots, corresponding to 28 different cutaneous proteins, were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and the Mascot online database searching algorithm. Twenty-five proteins were expressed at higher levels in the epidermis, whereas only nine were found predominantly in the subepidermal tissues. A subset of protein spots exhibited strain-specific expression. Proteins of diverse function were identified, including those involved in stress response, apoptosis, growth inhibition, the maintenance of structural integrity, translational control, energy metabolism, calcium binding, cholesterol transport, and the scavenging of free radicals. Prohibitin expression was detected cutaneously, with more abundant protein and mRNA levels in the epidermis. Five molecular chaperones including protein di-sulfide isomerase, 78 kDa glucose-regulated protein precursor, heat shock protein 60 (HSP60), HSP70, and HSP27 were also identified. Of these, HSP27 expression was confined mainly to the epidermis, and expression of protein disulfide isomerase was found primarily in the subepidermal tissues. Proteomic analysis of skin following heat or cold shock resulted in increased levels of HSP27, HSP60, and HSP70 suggesting involvement of these chaperones in the cutaneous response mechanism to temperature stress. These data establish numerous reference markers within the proteome map of murine skin and provide an important framework for future efforts aimed at characterization of the epidermal and subepidermal responses to environmental changes.