Improved structures of full-length p97, an AAA ATPase: Implications for mechanisms of nucleotide-dependent conformational change
Improved structures of full-length p97, an AAA ATPase: Implications for mechanisms of nucleotide-dependent conformational change
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DOI：
10.1016/j.str.2008.02.010
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发表时间：
2008-05-01
期刊：
影响因子：
5.7
通讯作者：
Weis, William I.
中科院分区：
文献类型：
--
作者：
Davies, Jason M.;Brunger, Axel T.;Weis, William I.
与各种细胞活动 (AAA) 蛋白 p97 相关的 ATP 酶与多种细胞过程有关，包括内质网相关降解和同型膜融合。 p97 属于 AAA 蛋白亚组，包含两个核苷酸结合域 D1 和 D2。我们以 3.0 埃的分辨率确定了 D2 的晶体结构。该模型能够根据之前报道的低分辨率衍射数据对不同核苷酸状态下的全长 p97 进行重新细化，从而显着改善 R 值和 Ramachandran 统计数据。尽管总体倍数保持相似，但仍有显着改进，尤其是在 D2 核苷酸结合位点周围。重新细化说明了了解覆盖整个分子大部分的片段高分辨率结构的重要性。这些结构表明，通过六聚体中相邻的一个 D2 结构域推动一个 D2 结构域，并通过 D1-D2 连接子传输核苷酸状态信息以取代 N 末端效应器结合结构域，核苷酸水解转化为更大的构象变化。
The ATPases associated with various cellular activities (AAA) protein p97 has been implicated in a variety of cellular processes, including endoplasmic reticulum-associated degradation and homotypic membrane fusion. p97 belongs to a subgroup of AAA proteins that contains two nucleotide binding domains, D1 and D2. We determined the crystal structure of D2 at 3.0 angstrom resolution. This model enabled rerefinement of full-length p97 in different nucleotide states against previously reported low-resolution diffraction data to significantly improved R values and Ramachandran statistics. Although the overall fold remained similar, there are significant improvements, especially around the D2 nucleotide binding site. The rerefinement illustrates the importance of knowledge of high-resolution structures of fragments covering most of the whole molecule. The structures suggest that nucleotide hydrolysis is transformed into larger conformational changes by pushing of one D2 domain by its neighbor in the hexamer, and transmission of nucleotide-state information through the D1-D2 linker to displace the N-terminal, effector binding domain.