Celis J E, Gesser B, Rasmussen H H, Madsen P, Leffers H, Dejgaard K, Honore B, Olsen E, Ratz G, Lauridsen J B
Institute of Medical Biochemistry, Aarhus University, Denmark.
Electrophoresis. 1990 Dec;11(12):989-1071. doi: 10.1002/elps.1150111202.
A total of 3430 polypeptides (2592 cellular; 838 secreted) from transformed human amnion cells (AMA) labeled with [35S]methionine were separated and recorded using computer-aided two-dimensional (2-D) gel electrophoresis. A master 2-D gel database of cellular protein information that includes both qualitative and quantitative annotations has been established. The protein numbers in this database differ from those reported in an earlier version (Celis et al. Leukemia 1988, 2,561-602) as a result of changes in the scanning hardware. The reported information includes: percentage of total radioactivity recovered from the gels (based on quantitations of polypeptides labeled with a mixture of 16 14C-amino acids), protein name (including credit to investigators that aided identification), antibody against protein, cellular localization, (nuclear, 40S hnRNP, 20S snRNP U5, proteasomes, endoplasmic reticulum, mitochondria, Golgi, ribosomes, intermediate filaments, microfilaments and microtubules), levels in fetal human tissues, partial protein sequences (containing information on 48 human proteins microsequenced so far), cell cycle-regulated proteins, proteins sensitive to interferons alpha, beta, and gamma, heat shock proteins, annexins and phosphorylated proteins. The results presented should be considered as the initial phase of a joint effort between our laboratories to undertake a general and systematic analysis of human proteins. Using this integrated approach it will be possible to identify phenotype-specific proteins, to microsequence them and store the information in the database, to identify the corresponding genes, to search for homology with previously characterized proteins and to study the function of groups of proteins (pathways, organelles, etc.) that exhibit interesting regulatory properties. In particular, the 2-D gel protein database may become increasingly important in view of the concerted effort to map and sequence the entire human genome.
用[35S]甲硫氨酸标记转化的人羊膜细胞(AMA),通过计算机辅助二维(2-D)凝胶电泳分离并记录了总共3430种多肽(2592种细胞内多肽;838种分泌型多肽)。已建立了一个包含定性和定量注释的细胞蛋白质信息二维凝胶主数据库。由于扫描硬件的变化,该数据库中的蛋白质数量与早期版本(Celis等人,《白血病》1988年,2,561 - 602)中报告的不同。报告的信息包括:从凝胶中回收的总放射性百分比(基于用16种14C - 氨基酸混合物标记的多肽的定量)、蛋白质名称(包括对协助鉴定的研究人员的致谢)、针对该蛋白质的抗体、细胞定位(细胞核、40S hnRNP、20S snRNP U5、蛋白酶体、内质网、线粒体、高尔基体、核糖体、中间丝、微丝和微管)、在人胎儿组织中的水平、部分蛋白质序列(包含迄今为止对48种人类蛋白质进行微测序的信息)、细胞周期调节蛋白、对α、β和γ干扰素敏感的蛋白质、热休克蛋白、膜联蛋白和磷酸化蛋白。所呈现的结果应被视为我们实验室之间共同努力对人类蛋白质进行全面系统分析的初始阶段。使用这种综合方法,将有可能鉴定表型特异性蛋白质、对其进行微测序并将信息存储在数据库中、鉴定相应基因、搜索与先前表征的蛋白质的同源性以及研究具有有趣调节特性的蛋白质组(途径、细胞器等)的功能。特别是,鉴于绘制和测序整个人类基因组的协同努力,二维凝胶蛋白质数据库可能会变得越来越重要。