Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology
Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology
复制标题
DOI：
10.1073/pnas.160270797
复制
发表时间：
2000-08-15
影响因子：
11.1
通讯作者：
Aebersold, R
中科院分区：
文献类型：
--
作者：
Gygi, SP;Corthals, GL;Aebersold, R
蛋白质组分析最常用的方法是结合双向凝胶电泳法(2DE)和质谱仪(MS)进行蛋白质分离和鉴定。尽管这项技术强大、成熟和灵敏，但关于其表征蛋白质组所有元素的能力仍然存在疑问。在目前的研究中，通过银染在狭窄的pH范围(4.9-5.7)2D凝胶中分离出0.5 mg的总可溶性酵母蛋白，显示了1500多个特征。对迁移到4 cm(2)区域的50个斑点进行了MS蛋白质鉴定。尽管样品载量很高，并扩大了电泳分离，但来自密码子偏好值基因的蛋白质
Proteome analysis is most commonly accomplished by a combination of two-dimensional gel electrophoresis (2DE) to separate and visualize proteins and mass spectrometry (MS) for protein identification. Although this technique is powerful, mature, and sensitive, questions remain concerning its ability to characterize all of the elements of a proteome. In the current study, more than 1,500 features were visualized by silver staining a narrow pH range (4.9-5.7) 2D gel in which 0.5 mg of total soluble yeast protein was separated. Fifty spots migrating to a region of 4 cm(2) were subjected to MS protein identification. Despite the high sample load and extended electrophoretic separation, proteins from genes with codon bias values of