皮肤细胞蛋白质组质谱分析方法的建立及肽质量指纹图谱的构建

Establishment of Mass Spectrometer and Peptide Mass Fingerprinting for Proteomics of Skin Cells

目的建立人皮肤细胞蛋白质组学研究所需的质谱分析方法,并获得皮肤细胞蛋白质肽质量指纹图谱。方法体外培养人皮肤成纤维细胞与角质形成细胞,采用固相pH梯度双向凝胶电泳分离其总蛋白质,凝胶经染色后,在图谱上选取2个角质形成细胞蛋白点与1个成纤维细胞蛋白点,进行胶内酶切、基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析鉴定。结果获得3个点的肽质量指纹图谱,经Mascot软件搜索人非冗余蛋白质数据库后,鉴定为细胞骨架Actingamma1、tubulinα2及热休克蛋白HSP70。结论皮肤细胞基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析技术的建立为皮肤蛋白质组学的进一步研究提供手段。

Objective Preliminary establish of mass spectrometer and peptide mass fingerprinting for the proteome research of skin cells. Methods The total proteins of fibroblast and keratinocyte were separated by immobilized pH gradient based two dimensional gel electmphoresis （2 - DE）. After staining, Matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS） were performed with 2 proteins from keratinocyte and 1 proteins from fibroblast. Results 3 Peptide Mass Fingerprintings were obtained by MALDI-TOF-MS, and 3 protein spots were identified by using the Mascot soft ware. They were Actin gamma 1,tubulin α2 and HSP 70 kDa. Conclusion The establishment of MALDI-TOF-MS and peptide mass fingerprinting will be helpful in the further proteomic study of skin.