Ramaekers F C, Hukkelhoven M W, Groeneveld A, Bloemendal H
Biochim Biophys Acta. 1984 Jun 29;799(3):221-9. doi: 10.1016/0304-4165(84)90264-2.
Changes in biosynthesis of lens proteins upon culturing have been studied by one- and two-dimensional gel electrophoretic techniques. In primary cells still growing on the capsule, alpha B2-crystallin is synthesized in a relatively high amount next to the main cytoskeletal constituents actin, tubulin and vimentin. In addition, a minor amount of beta Bp seems to be synthesized too. When the cells grow off the capsule, alpha-crystallin synthesis diminishes. beta-Crystallin synthesis continues at a low rate in cells growing on plastic or in cells forming 'lentoid bodies'. When the cells are subcultured, the synthesis of actin and vimentin becomes more pronounced, while tubulin synthesis is no longer detectable after three transfers. The relative amount of vimentin decreases, as compared to actin, during aging and elongation of the cells. When the cells have been transferred ten times and have started to elongate, a 55 kDa protein doublet differing from tubulin is observed in the two-dimensional gel patterns. We observed that elongation of lens cells in culture is accompanied by an increase in the synthesis of a polypeptide of the 26 kDa region. Furthermore, a major glycoprotein is found in the 130 kDa region, but overall glycosylation of proteins seems to decrease during lens cell elongation in vitro.
通过一维和二维凝胶电泳技术研究了培养过程中晶状体蛋白生物合成的变化。在仍在晶状体囊膜上生长的原代细胞中，除了主要的细胞骨架成分肌动蛋白、微管蛋白和波形蛋白外，αB2-晶状体蛋白的合成量相对较高。此外，似乎也合成少量的βBp。当细胞从囊膜上脱离生长时，α-晶状体蛋白的合成减少。在塑料上生长的细胞或形成“类晶状体小体”的细胞中，β-晶状体蛋白的合成以低速率持续进行。当细胞进行传代培养时，肌动蛋白和波形蛋白的合成变得更加明显，而在三次传代后微管蛋白的合成不再能检测到。在细胞老化和伸长过程中，与肌动蛋白相比，波形蛋白的相对量减少。当细胞传代十次并开始伸长时，在二维凝胶图谱中观察到一种与微管蛋白不同的55 kDa蛋白双峰。我们观察到培养的晶状体细胞伸长伴随着26 kDa区域一种多肽合成的增加。此外，在130 kDa区域发现一种主要的糖蛋白，但在体外晶状体细胞伸长过程中蛋白质的总体糖基化似乎减少。
