SELENIUM & BROMINE DOUBLE EDGE MAD EXPERIMENT FOR AN HSF DNA COMPLEX

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• 批准号: 6586596
• 负责人: HILLARY C NELSON
• 金额: $ 14.32万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6586596
• 关键词: bioimaging /biomedical imaging; biomaterials; biomedical resource; genome; proteins; radiography; structural biology

We have used multiple anomalous disperson (MAD) to determine the structure of a complex between the DNA binding domain of the heat shock transcription factor (HSF) along with a 12-mer of DNA containing its cognate binding site. Data was collected on crystals containing selenomethionine-substituted protein and bromouracil-substituted DNA. The present model contains complete DNA, most of the protein, and 100 water molecules. The protein monomers are missing residues from the N- and C-terminii, as well as from the flexible loop. The current R value is 21.0% (free R is 29.3%) for all data between 6.0 and 2.5 E. There are no Ramachandran outliers, the r.m.s deviations are 0.009 E for bond lengths and 1.5540 for bond angles. The structure shows that the DNA is contacted solely along with phosphate backbone, with the majority of contacts coming from the putative recognition helix of the DNA binding domain. We are in the process of interpreting the biological implications of the structure.

我们使用多重异常驱散（MAD）来确定 热 DNA 结合结构域之间的复合物结构 休克转录因子 (HSF) 以及含有 12 聚体 DNA 它的同源结合位点。 收集的数据包含晶体 硒代蛋氨酸取代的蛋白质和溴尿嘧啶取代的DNA。 目前的模型包含完整的 DNA、大部分蛋白质和 100 水分子。 蛋白质单体缺少残基 N 端和 C 端，以及来自柔性环。 当前R 对于 6.0 到 2.5 E 之间的所有数据，值为 21.0%（自由 R 为 29.3%）。 没有 Ramachandran 异常值，均方根偏差为 0.009 E 键长为 1.5540，键角为 1.5540。 结构表明 DNA 仅与磷酸骨架接触， 大多数接触来自假定的识别螺旋 DNA 结合域。 我们正在解释 该结构的生物学意义。