Lenstra J A, Bloemendal H
Eur J Biochem. 1983 Oct 3;135(3):413-23. doi: 10.1111/j.1432-1033.1983.tb07668.x.
Chemical extractions are proposed as a major tool for a fractionation of cellular proteins. As a model system, proteins from cultured hamster lens cells have been divided by independent extractions into seven subcellular fractions, corresponding to water-soluble proteins and the proteins from membranes, microfilaments (and other deoxycholate-soluble proteins), intermediate filaments, microtubules, polysomes and nuclei respectively. The latter two fractions have been subfractionated yielding ribosomal proteins, the elongation and initiation factors of the protein-synthesis machinery, chromatin proteins and non-chromatin proteins. The protein compositions of the fractions have been analyzed by one-dimensional and two-dimensional gel electrophoresis. This resulted in an almost complete topography of the proteins detected on two-dimensional gels of total-cell lysates. Comparison of two-dimensional patterns of proteins from the total-cell lysate and proteins from hamster erythrocytes or from liver, muscle or brain tissue showed that the different cell types have only few proteins in common. Two proteins are common to all of these cell types, namely actin and a 68-kDa protein. The latter protein was, like actin, vimentin and the tubulin subunits, also present in most cell fractions. Evidence is presented that this protein is identical to a 68-kDa heat-shock protein.
化学提取法被认为是细胞蛋白质分级分离的主要工具。作为一个模型系统，培养的仓鼠晶状体细胞中的蛋白质通过独立提取被分为七个亚细胞组分，分别对应水溶性蛋白质以及来自膜、微丝（和其他脱氧胆酸盐可溶蛋白质）、中间丝、微管、多核糖体和细胞核的蛋白质。后两个组分已被进一步细分，得到核糖体蛋白质、蛋白质合成机制的延伸因子和起始因子、染色质蛋白质和非染色质蛋白质。通过一维和二维凝胶电泳分析了这些组分的蛋白质组成。这几乎得到了全细胞裂解物二维凝胶上检测到的蛋白质的完整图谱。全细胞裂解物中的蛋白质与仓鼠红细胞、肝脏、肌肉或脑组织中的蛋白质的二维图谱比较表明，不同细胞类型只有很少的蛋白质是共有的。所有这些细胞类型共有两种蛋白质，即肌动蛋白和一种68 kDa的蛋白质。后一种蛋白质与肌动蛋白、波形蛋白和微管蛋白亚基一样，也存在于大多数细胞组分中。有证据表明这种蛋白质与一种68 kDa的热休克蛋白相同。
