SELENOMETHIONYL PROTEINS PRODUCED FOR ANALYSIS BY MULTIWAVELENGTH ANOMALOUS DIFFRACTION (MAD) - A VEHICLE FOR DIRECT DETERMINATION OF 3-DIMENSIONAL STRUCTURE
SELENOMETHIONYL PROTEINS PRODUCED FOR ANALYSIS BY MULTIWAVELENGTH ANOMALOUS DIFFRACTION (MAD) - A VEHICLE FOR DIRECT DETERMINATION OF 3-DIMENSIONAL STRUCTURE
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DOI：
10.1002/j.1460-2075.1990.tb08287.x
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发表时间：
1990-05-01
期刊：
影响因子：
11.4
通讯作者：
LEMASTER, DM
中科院分区：
文献类型：
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作者：
HENDRICKSON, WA;HORTON, JR;LEMASTER, DM
已建立了一个表达系统，用于将硒代蛋氨酸掺入大肠杆菌质粒产生的重组蛋白中。硒代蛋氨酸对T4和E的替代率均为100%。大肠杆菌硫氧还蛋白天然的重组蛋白质和两种硫氧还蛋白的硒代甲硫酰变体同晶结晶。异常散射因子推导出从同步X射线吸收测量的硒甲硫酰蛋白质的晶体。采取参考的经验，硒生物素链霉亲和素的结构分析的方法，多波长异常衍射（MAD），这些数据表明，重组硒蛋氨酸蛋白质分析MAD定相提供了一个相当普遍的手段，阐明原子结构。
An expression system has been established for the incorporation of selenomethionine into recombinant proteins produced from plasmids in Escherichia coli. Replacement of methionine by selenomethionine is demonstrated at the level of 100% for both T4 and E. coli thioredoxins. The natural recombinant proteins and the selenomethionyl variants of both thioredoxins crystallize isomorphously. Anomalous scattering factors were deduced from synchroton X-ray absorption measurements of crystals of the selenomethionyl proteins. Taken with reference to experience in the structural analysis of selenobiotinyl streptavidin by the method of multiwavelength anomalous diffraction (MAD), these data indicate that recombinant selenomethionyl proteins analyzed by MAD phasing offer a rather general means for the elucidation of atomic structures.