Huang Chun-Ming, Foster K Wade, DeSilva Tivanka, Zhang JianFeng, Shi Zhongkai, Yusuf Nabiha, Van Kampen Kent R, Elmets Craig A, Tang De-chu C
Department of Dermatology, University of Alabama at Birmingham, USA.
J Invest Dermatol. 2003 Jul;121(1):51-64. doi: 10.1046/j.1523-1747.2003.12327.x.
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.
哺乳动物的皮肤经常受到不同环境压力的影响，每种压力都会导致蛋白质表达发生特定的补偿性变化，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谱，在单一的二维聚丙烯酰胺凝胶中可以分辨出500多个蛋白质斑点。使用基质辅助激光解吸/电离飞行时间质谱和Mascot在线数据库搜索算法，鉴定出了44个蛋白质斑点，对应于28种不同的皮肤蛋白质。25种蛋白质在表皮中表达水平较高，而只有9种主要在表皮下组织中发现。一部分蛋白质斑点表现出菌株特异性表达。鉴定出了功能多样的蛋白质，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。在皮肤中检测到了抑制素的表达，在表皮中的蛋白质和mRNA水平更为丰富。还鉴定出了5种分子伴侣，包括蛋白质二硫键异构酶、78 kDa葡萄糖调节蛋白前体、热休克蛋白60（HSP60）、HSP70和HSP27。其中，HSP27的表达主要局限于表皮，而蛋白质二硫键异构酶的表达主要在表皮下组织中发现。对热休克或冷休克后的皮肤进行蛋白质组学分析，结果显示HSP27、HSP60和HSP70的水平升高，表明这些分子伴侣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谱中建立了众多参考标记，并为未来旨在表征表皮和表皮下对环境变化反应的研究提供了重要框架。
