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Protein crystallization screening using enhanced associative experimental design

机译:使用增强的联想实验设计蛋白质结晶筛选

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Protein crystallization screening helps determine factors (e.g., salts, pH of buffers, ionic strengths, temperature, and type of precipitants) that are favorable for the formation of a large protein crystal suitable for X-ray crystallography. While existing commercial screens may not generate crystalline outcomes for difficult proteins, their outcomes could be used for recommending novel screens. Current methods for protein crystallization screening such as associative experimental design (AED) process only cocktails having one chemical per reagent while ignoring cocktails with multiple chemicals per reagent. To analyze cocktails having multiple chemicals per reagent, we propose enhanced associative experimental design (AED) that recommends novel crystallization conditions by analyzing the content of successful preliminary crystallization conditions. In wet lab experiments, our enhanced AED (AED~+) yielded ten new crystalline conditions for Tt189 (Nucleoside diphosphate kinase) in addition to 20 crystalline conditions generated by AED. Moreover, our AED~+ allows pairing of crystalline or likely lead outcome with a non-crystalline outcome to generate novel crystalline conditions overcoming the limitation of AED requiring at least two good cocktails having at least one coming reagent.
机译:蛋白质结晶筛选有助于确定因素(例如,缓冲剂,离子强度,沉淀剂的沉淀物的盐,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,温度,沉淀物的类型)是有利的,这对于适于形成适用于X射线晶体学的大蛋白质晶体。虽然现有的商业屏幕可能不会产生困难蛋白质的结晶结果,但它们的结果可用于推荐新颖的屏幕。目前蛋白质结晶筛选的方法如关联实验设计(AED)过程仅在每种试剂中具有一个化学品的鸡尾酒,同时忽略每种试剂多种化学品的鸡尾酒。为了分析每种试剂具有多种化学品的鸡尾酒,我们通过分析成功初步结晶条件的含量提高了增强的联想实验设计(AED),推荐新的结晶条件。在湿实验室实验中,我们的增强型AED(AED〜+)除了通过AED产生的20个晶体条件外,还产生了TT189(核苷二磷酸激酶)的10个新的结晶条件。此外,我们的AED〜+允许与非结晶结果配对结晶或可能的铅结果,以产生新的晶体条件克服AED的限制,要求至少有两个具有至少一种即将到来的试剂的良好鸡尾酒。

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