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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Shotgun crystallization strategy for structural genomics: an optimized two-tiered crystallization screen against the Thermotoga maritima proteome.
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Shotgun crystallization strategy for structural genomics: an optimized two-tiered crystallization screen against the Thermotoga maritima proteome.

机译：猎枪结晶结构的策略基因组学:一个优化的层的结晶屏幕对Thermotoga maritima蛋白质组。

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摘要

As the field of structural genomics continues to grow and new technologies are developed, novel strategies are needed to efficiently crystallize large numbers of protein targets, thus increasing output, not just throughput [Chayen & Saridakis (2002). Acta Cryst. D58, 921-927]. One strategy, developed for the high-throughput structure determination of the Thermotoga maritima proteome, is to quickly determine which proteins have a propensity for crystal formation followed by focused SeMet-incorporated protein crystallization attempts. This experimental effort has resulted in over 320 000 individual crystallization experiments. As such, it has provided one of the most extensive systematic data sets of commonly used crystallization conditions against a wide range of proteins to date. Analysis of this data shows that many of the original screening conditions are redundant, as all of the T. maritima proteins that crystallize readily could be identified using just 23% of the original conditions. It also showsthat proteins that contain selenomethionine and are more extensively purified often crystallize in distinctly different conditions from those of their native less pure counterparts. Most importantly, it shows that the two-tiered strategy employed here is extremely successful for predicting which proteins will readily crystallize, as greater than 99% of the proteins identified as having a propensity to crystallize under non-optimal native conditions did so again as selenomethionine derivatives during the focused crystallization trials. This crystallization strategy can be adopted for both large-scale genomics programs and individual protein studies with multiple constructs and has the potential to significantly accelerate future crystallographic efforts.
机译：结构基因组学领域的继续成长和新技术开发、小说策略需要有效的结晶大量的蛋白质的目标,从而增加输出,不仅吞吐量(Chayen & Saridakis(2002)。为高通量开发结构确定Thermotoga maritima蛋白质组,迅速确定哪些蛋白质有一个倾向晶体形成后通过集中SeMet-incorporated蛋白结晶的尝试。努力导致320多000人结晶实验。提供的一个最广泛的系统数据集的常用的结晶条件对多种蛋白质日期。最初的筛选条件是多余的,所有的t . maritima蛋白质结晶很容易可以确定使用只有23%的原始条件。表明蛋白质含有硒代蛋氨酸经常和更广泛的纯化在截然不同的条件下结晶与本国的不纯同行。这里采用双重策略非常成功的预测蛋白质容易结晶,大于99%的蛋白质鉴定为倾向没有优化的本地条件下结晶所以作为硒代蛋氨酸衍生品吗在集中结晶试验。结晶策略既可以采用大规模基因组学项目和个人蛋白质与多个构造和研究未来的潜力大大加速晶体的努力。

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