A Docking Model Based on Mass Spectrometric and Biochemical Data Describes Phage Packaging Motor Incorporation
A Docking Model Based on Mass Spectrometric and Biochemical Data Describes Phage Packaging Motor Incorporation
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DOI：
10.1074/mcp.m900625-mcp200
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发表时间：
2010-08-01
影响因子：
7
通讯作者：
Prevelige, Peter E., Jr.
中科院分区：
文献类型：
--
作者：
Fu, Chi-yu;Uetrecht, Charlotte;Prevelige, Peter E., Jr.
由于缺乏连接器和脚手架蛋白如何相互作用的结构信息，在羔羊样噬菌体组装过程中，脚手架蛋白介导的一个且仅一个DNA包装马达/连接器十二聚体在一个独特顶点的结合的分子机制仍然是难以捉摸的。我们组装并表征了一个phi 29连接-支架复合物，该复合物可以在体外组装过程中并入原囊体。原生质谱分析显示，该连接器结合最多12个支架分子，可能组织为6个二聚体。一个数据驱动的对接模型，使用化学交联和诱变数据的输入，表明脚手架蛋白和连接十二聚体宽结构域的外部之间存在相互作用。支架蛋白的接头结合区位于C端衣壳结合区上游。这种排列允许复合物内支架蛋白的C端招募衣壳亚基并介导单个连接顶点的结合。中国生物医学工程学报（英文版），2010。
The molecular mechanism of scaffolding protein-mediated incorporation of one and only one DNA packaging motor/connector dodecamer at a unique vertex during lambdoid phage assembly has remained elusive because of the lack of structural information on how the connector and scaffolding proteins interact. We assembled and characterized a phi 29 connector-scaffolding complex, which can be incorporated into procapsids during in vitro assembly. Native mass spectrometry revealed that the connector binds at most 12 scaffolding molecules, likely organized as six dimers. A data-driven docking model, using input from chemical cross-linking and mutagenesis data, suggested an interaction between the scaffolding protein and the exterior of the wide domain of the connector dodecamer. The connector binding region of the scaffolding protein lies upstream of the capsid binding region located at the C terminus. This arrangement allows the C terminus of scaffolding protein within the complex to both recruit capsid subunits and mediate the incorporation of the single connector vertex. Molecular & Cellular Proteomics 9:1764-1773, 2010.