Beuron Fabienne, Dreveny Ingrid, Yuan Xuemei, Pye Valerie E, McKeown Ciaran, Briggs Louise C, Cliff Matthew J, Kaneko Yayoi, Wallis Russell, Isaacson Rivka L, Ladbury John E, Matthews Steve J, Kondo Hisao, Zhang Xiaodong, Freemont Paul S
Centre for Structural Biology, Division of Molecular Biosciences, Imperial College London, South Kensington, UK.
EMBO J. 2006 May 3;25(9):1967-76. doi: 10.1038/sj.emboj.7601055. Epub 2006 Apr 6.
The AAA+ATPase p97/VCP, helped by adaptor proteins, exerts its essential role in cellular events such as endoplasmic reticulum-associated protein degradation or the reassembly of Golgi, ER and the nuclear envelope after mitosis. Here, we report the three-dimensional cryo-electron microscopy structures at approximately 20 Angstroms resolution in two nucleotide states of the endogenous hexameric p97 in complex with a recombinant p47 trimer, one of the major p97 adaptor proteins involved in membrane fusion. Depending on the nucleotide state, we observe the p47 trimer to be in two distinct arrangements on top of the p97 hexamer. By combining the EM data with NMR and other biophysical measurements, we propose a model of ATP-dependent p97(N) domain motions that lead to a rearrangement of p47 domains, which could result in the disassembly of target protein complexes.
AAA+ATP酶p97/VCP在衔接蛋白的帮助下，在细胞活动中发挥重要作用，如内质网相关蛋白降解或有丝分裂后高尔基体、内质网和核膜的重新组装。在此，我们报告了内源性六聚体p97与重组p47三聚体（参与膜融合的主要p97衔接蛋白之一）形成复合物在两种核苷酸状态下约20埃分辨率的三维冷冻电子显微镜结构。根据核苷酸状态，我们观察到p47三聚体在p97六聚体顶部有两种不同的排列方式。通过将电子显微镜数据与核磁共振及其他生物物理测量相结合，我们提出了一个ATP依赖的p97(N)结构域运动模型，该模型导致p47结构域重排，这可能导致靶蛋白复合物的解体。
