ATP binding to p97/VCP D1 domain regulates selective recruitment of adaptors to its proximal N-domain.
ATP binding to p97/VCP D1 domain regulates selective recruitment of adaptors to its proximal N-domain.
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DOI：
10.1371/journal.pone.0050490
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Geifman Shochat S
中科院分区：
文献类型：
--
作者：
Chia WS;Chia DX;Rao F;Bar Nun S;Geifman Shochat S
p97/Valosin-containing protein（VCP）是AAA-ATP酶家族的一员，参与细胞分裂、胞内运输和内质网相关降解（ERAD）中错误折叠蛋白的提取等多种细胞过程。它是一种同型六聚体，每个亚基含有两个串联的D1和D2 ATP酶结构域以及作为接头蛋白结合结构域的N-和C-末端区域。p97/VCP通过与各种接头蛋白结合而导向其许多不同的功能途径。衔接蛋白的募集的调节仍不清楚。两个衔接蛋白，Ufd 1/Np 14和p47，专门结合到p97/VCP N-结构域和直接p97/VCP的ERAD相关的过程或高尔基体片段的同型融合，在这里进行了研究。基于表面等离子体共振生物传感器的测定允许在真实的时间内研究结合动力学。在竞争实验中，观察到在ATP存在下，Ufd 1/Np 14能够更有效地与p47竞争结合p97/VCP。通过使用不可水解的ATP类似物和六聚体截短的p97/N-D1片段，表明ATP与近端D1结构域的结合而不是水解增强了Ufd 1/Np 14与N结构域的结合，从而调节Ufd 1/Np 14或p47的募集。ATP的这种新作用和D1 AAA-ATP酶结构域的指定功能将p97/VCP的多种功能与细胞的代谢状态联系起来。
p97/Valosin-containing protein (VCP) is a member of the AAA-ATPase family involved in many cellular processes including cell division, intracellular trafficking and extraction of misfolded proteins in endoplasmic reticulum-associated degradation (ERAD). It is a homohexamer with each subunit containing two tandem D1 and D2 ATPase domains and N- and C-terminal regions that function as adaptor protein binding domains. p97/VCP is directed to its many different functional pathways by associating with various adaptor proteins. The regulation of the recruitment of the adaptor proteins remains unclear. Two adaptor proteins, Ufd1/Npl4 and p47, which bind exclusively to the p97/VCP N-domain and direct p97/VCP to either ERAD-related processes or homotypic fusion of Golgi fragments, were studied here. Surface plasmon resonance biosensor-based assays allowed the study of binding kinetics in real time. In competition experiments, it was observed that in the presence of ATP, Ufd1/Npl4 was able to compete more effectively with p47 for binding to p97/VCP. By using non-hydrolysable ATP analogues and the hexameric truncated p97/N-D1 fragment, it was shown that binding rather than hydrolysis of ATP to the proximal D1 domain strengthened the Ufd1/Npl4 association with the N-domain, thus regulating the recruitment of either Ufd1/Npl4 or p47. This novel role of ATP and an assigned function to the D1 AAA-ATPase domain link the multiple functions of p97/VCP to the metabolic status of the cell.
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DOI：
10.1038/nsb972
发表时间：
2003-10-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
DeLaBarre, B;Brunger, AT
通讯作者：
Brunger, AT
影响因子：
5.6
作者：
Beuron, F;Flynn, TC;Freemont, PS
通讯作者：
Freemont, PS
影响因子：
3
作者：
Huyton, T;Pye, VE;Freemont, PS
通讯作者：
Freemont, PS
影响因子：
12.4
作者：
Hirabayashi, M;Inoue, K;Kakizuka, A
通讯作者：
Kakizuka, A
影响因子：
11.4
作者：
Braun, S;Matuschewski, K;Jentsch, S
通讯作者：
Jentsch, S