Pechkova Eugenia, Nicolini Claudio
Department of Biophysical M&O Sciences and Technologies-Genoa University, Genoa, Italy.
J Cell Biochem. 2002;85(2):243-51. doi: 10.1002/jcb.10123.
A new method of protein nucleation and crystallization based on Langmuir-Blodgett technology is here utilized for the template stimulation of crystal growth of so far non-crystallized proteins. Microcrystals (60-120 microm) of bovine cytochrome P450scc and human protein kinase CKII alpha subunit were obtained with use of the homologous protein thin film template by vapor diffusion modified hanging drop method. The induction of microcrystals nucleation by the thin template confirms in the two different important classes of proteins, until now never crystallized, the positive stimulatory influence for crystal formation of protein thin film template, which was observed in an earlier study with a model system (chicken egg white lysozyme) as an unexpected acceleration and enhancement in the crystal growth.
本文采用一种基于朗缪尔-布洛杰特技术的蛋白质成核与结晶新方法，用于对迄今尚未结晶的蛋白质晶体生长进行模板刺激。通过气相扩散改良悬滴法，利用同源蛋白质薄膜模板获得了牛细胞色素P450scc和人蛋白激酶CKIIα亚基的微晶（60 - 120微米）。薄模板对微晶成核的诱导在两类不同的重要蛋白质中得到证实，这两类蛋白质迄今从未结晶，蛋白质薄膜模板对晶体形成具有积极的刺激作用，这在早期以模型系统（鸡蛋白溶菌酶）进行的研究中被观察到，表现为晶体生长的意外加速和增强。
