Hynes G, Sutton C W, U S, Willison K R
CRC Centre for Cell and Molecular Biology, Institute of Cancer Research, Chester Beatty Laboratories, London, United Kingdom.
FASEB J. 1996 Jan;10(1):137-47. doi: 10.1096/fasebj.10.1.8566534.
The chaperonin-containing TCP-1 (CCT), found in the eukaryotic cytosol, is currently the focus of extensive research, CCT isolated from mouse testis lysate sediments at 20S in a sucrose gradient and accounts for about 70% of the total protein in this fraction. We intend to identify all the other proteins that copurify with CCT and to compile a reference profile for future studies. Their identification can be accelerated by a combination of protease digestion, matrix-assisted laser desorption-mass spectrometry, and database matching known as peptide mass fingerprinting. We applied this strategy to 32 polypeptides resolved by 2-dimensional gel electrophoresis, and 23 known proteins and 6 novel proteins were identified. We analyzed isoelectric variants of the CCT subunits and differences in the peptide mass spectra of two CCT theta isoforms indicated a novel posttranslational modification of this subunit.
存在于真核细胞质中的含伴侣蛋白TCP-1（CCT），目前是广泛研究的焦点。从鼠睾丸裂解物中分离出的CCT在蔗糖梯度中于20S处沉降，占该组分中总蛋白的约70%。我们打算鉴定与CCT共纯化的所有其他蛋白质，并编制一份参考图谱以供未来研究使用。通过蛋白酶消化、基质辅助激光解吸质谱以及称为肽质量指纹图谱的数据库匹配相结合的方法，可以加速对它们的鉴定。我们将此策略应用于通过二维凝胶电泳分离的32种多肽，鉴定出了23种已知蛋白质和6种新蛋白质。我们分析了CCT亚基的等电变体，两种CCTθ亚型的肽质量谱差异表明该亚基存在一种新的翻译后修饰。
