大鼠内耳表达蛋白质组的2-DE分离和初步鉴定

Separation and analysis of proteomics of the rat inner ear by 2-DE and MS

目的通过制备、分离和鉴定正常情况下内耳表达的蛋白质,了解内耳的蛋白质组学特征。方法本研究以正常大鼠内耳组织为起始材料,通过2-DE分离内耳表达的蛋白质组,利用MALDI-TOF质谱仪对分离的蛋白质进行分析和鉴定。结果从20只新生大鼠内耳组织提取到800μg蛋白质,在18cm×20cm的2-DE胶上可分离到300多个蛋白质斑点,质谱仪鉴定提示这些蛋白质分别属于细胞结构、细胞分裂、代谢相关、细胞信号传导、细胞因子相关蛋白质及未知蛋白质。结论本研究建立了大鼠内耳蛋白质组的分离和提取的方法,初步构建内耳2-DE参考图谱,并对其中的部分蛋白质进行了鉴定和分类,该图谱有助于内耳研究,尤其是不同状态下如疾病时内耳蛋白质表达变化的研究。

Objective To investigate proteomics of the inner ear by two direction electrophoresis and biologic mass spectrometry. Methods In this study we used the inner ear of newborn rats as the primary materials to conduct proteomics analysis of the ear by 2-DE and MS. Results In an 18 cm ~ 20 cm 2-D gel we separated more than 300 protein dots from inner ear tissues after cut and digested. These proteins were analyzed by MALDI-TOF MS, and the results underwent processing. Conclusion We established a method for sample collection and separation of the proteome of the inner ear in 2-DE gel, and also constructed a primary reference 2-DE map of the rat inner ear.