Christendat D, Yee A, Dharamsi A, Kluger Y, Savchenko A, Cort J R, Booth V, Mackereth C D, Saridakis V, Ekiel I, Kozlov G, Maxwell K L, Wu N, McIntosh L P, Gehring K, Kennedy M A, Davidson A R, Pai E F, Gerstein M, Edwards A M, Arrowsmith C H
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto 610 University Ave, Toronto, Ontario, Canada M5G 2M9.
Nat Struct Biol. 2000 Oct;7(10):903-9. doi: 10.1038/82823.
A set of 424 nonmembrane proteins from Methanobacterium thermoautotrophicum were cloned, expressed and purified for structural studies. Of these, approximately 20% were found to be suitable candidates for X-ray crystallographic or NMR spectroscopic analysis without further optimization of conditions, providing an estimate of the number of the most accessible structural targets in the proteome. A retrospective analysis of the experimental behavior of these proteins suggested some simple relations between sequence and solubility, implying that data bases of protein properties will be useful in optimizing high throughput strategies. Of the first 10 structures determined, several provided clues to biochemical functions that were not detectable from sequence analysis, and in many cases these putative functions could be readily confirmed by biochemical methods. This demonstrates that structural proteomics is feasible and can play a central role in functional genomics.
克隆、表达并纯化了嗜热自养甲烷杆菌的一组424种非膜蛋白，用于结构研究。其中，约20%被发现是X射线晶体学或核磁共振光谱分析的合适候选对象，无需进一步优化条件，这为蛋白质组中最易解析的结构靶点数量提供了一个估计。对这些蛋白质实验行为的回顾性分析表明，序列与溶解度之间存在一些简单关系，这意味着蛋白质特性数据库将有助于优化高通量策略。在前10个确定的结构中，有几个为序列分析无法检测到的生化功能提供了线索，而且在许多情况下，这些假定的功能可以通过生化方法轻易得到证实。这表明结构蛋白质组学是可行的，并且可以在功能基因组学中发挥核心作用。
