CO CRYSTALS OF HUMAN REPLICATION PROTEIN A & SINGLE STRANDED DNA

复制蛋白A(RPA)是DNA的关键组成部分 复制和修复机器。RPA是单链DNA 与DNA复制相互作用的结合蛋白 蛋白质、DNA聚合酶、转录激活蛋白和DNA 修复因子，包括那些在几种形式的 人类癌症。RPA是一种杂三聚体；DNA结合域已经被 定位到最大的亚单位。我们使用了有限的蛋白水解法 和缺失分析鉴定最小单链DNA 结合结构域。我们已经和单链DNA一起结晶了这个结构域。 受低温保护的晶体在我们的 RAXIS-II配备了双双对焦镜。我们是 正在对我们的部队进行重原子搜索，并准备好 硒蛋氨酸标记的蛋白质共结晶。在这 建议，我们希望使用，收集更高的分辨率原生以及 重原子数据集。

Replication protein A (RPA) is a key component of the DNA replication and repair machineries. RPA is a single stranded DNA binding protein that interacts with DNA replication orgin binding proteins, DNA polymerases, transcription activator proteins and DNA repair factors, including those that are mutated in several forms of human cancer. RPA is a heterotrimer; the DNA binding domain has been localized to the largest subunit. We have used limited proteolysis and deletion analysis to identify the minimal single strand DNA binding domain. We have cryslalized this domain together with ssDNA. Cryo-protected crystals diffract to a resolution of 2.2A on our RAXIS-II equipped with double double focussing mirrors. We are pursuing heavy atom searches on our unit and have prepared selenomethionine-labelled protein for co-crystallization. In this proposal, we wish to use, collect higher resolution native as well as heavy atom data sets.