Academic Journal

Lessons from ten years of crystallization experiments at the SGC.

التفاصيل البيبلوغرافية
العنوان: Lessons from ten years of crystallization experiments at the SGC.
المؤلفون: Ng, Jia Tsing, Dekker, Carien, Reardon, Paul, von Delft, Frank
المصدر: Acta Crystallographica: Section D, Structural Biology; Feb2016, Vol. 72 Issue 2, p224-235, 11p
مصطلحات موضوعية: CRYSTALLIZATION, RETROSPECTIVE studies, STRUCTURAL genomics
مستخلص: Although protein crystallization is generally considered more art than science and remains significantly trial-and-error, large-scale data sets hold the promise of providing general learning. Observations are presented here from retrospective analyses of the strategies actively deployed for the extensive crystallization experiments at the Oxford site of the Structural Genomics Consortium (SGC), where comprehensive annotations by SGC scientists were recorded on a customized database infrastructure. The results point to the importance of using redundancy in crystallizing conditions, specifically by varying the mixing ratios of protein sample and precipitant, as well as incubation temperatures. No meaningful difference in performance could be identified between the four most widely used sparse-matrix screens, judged by the yield of crystals leading to deposited structures; this suggests that in general any comparison of screens will be meaningless without extensive cross-testing. Where protein sample is limiting, exploring more conditions has a higher likelihood of being informative by yielding hits than does redundancy of either mixing ratio or temperature. Finally, on the logistical question of how long experiments should be stored, 98% of all crystals that led to deposited structures appeared within 30 days. Overall, these analyses serve as practical guidelines for the design of initial screening experiments for new crystallization targets. [ABSTRACT FROM AUTHOR]
Copyright of Acta Crystallographica: Section D, Structural Biology is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
قاعدة البيانات: Complementary Index
الوصف
تدمد:09074449
DOI:10.1107/S2059798315024687