Crystal structure of the MYB domain of the RAD transcription factor from Antirrhinum majus
Crystal structure of the MYB domain of the RAD transcription factor from Antirrhinum majus
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DOI：
10.1002/prot.21136
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发表时间：
2006-12-01
影响因子：
2.9
通讯作者：
Lawson, David M.
中科院分区：
文献类型：
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作者：
Stevenson, Clare E. M.;Burton, Nicolas;Lawson, David M.
材料与方法。详细介绍了A. majus RAD蛋白的制备以及8 kDa蛋白水解片段的制备和结晶已经在前面描述过。简言之，通过在100 mM CHES缓冲液（pH 9.5）中的2.8 M硫酸铵中进行气相扩散，使包含93个残基的天然RAD序列的残基6-74的片段结晶。晶体符合空间群P41212（或P43212），近似晶胞参数为a <$B $45 A和c <$72 A。这给出了晶体堆积参数（VM）为2.3 A 3 Daq 11，对应于不对称单元中片段的单个拷贝的估计溶剂含量为46%。对于低温数据收集，晶体需要用渗透压匹配的0.52 M硫酸铵溶液进行仔细的低温保护。
Materials and Methods. The detailed procedures for the overproduction of the A. majus RAD protein and the preparation and crystallization of the 8 kDa proteolytic fragment have been described previously. 18 Briefly, a fragment comprising residues 6–74 of the 93-residue native RAD sequence was crystallized by vapor diffusion from 2.8 M ammonium sulphate in 100 mM CHES buffer pH 9.5. Crystals conformed to space group P41212 (or P43212), with approximate cell parameters of a ¼ b ¼ 45 Aand c ¼ 72 A. This gave a crystal packing parameter (VM) of 2.3 A 3 DaÀ1 corresponding to an estimated solvent content of 46% for a single copy of the fragment in the asymmetric unit. For cryogenic data collection, the crystals required careful cryoprotection with an osmolality-matched19 solution of 0.52 M ammonium sul-