NAD(H) phosphates mediate tetramer assembly of human C-terminal binding protein (CtBP).
NAD(H) phosphates mediate tetramer assembly of human C-terminal binding protein (CtBP).
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DOI：
10.1016/j.jbc.2021.100351
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Royer WE Jr
中科院分区：
文献类型：
--
作者：
Nichols JC;Schiffer CA;Royer WE Jr
C-末端结合蛋白（CtBPs）是一类在细胞命运中起关键作用的共转录因子。我们先前已经表明NAD（H）促进来自人CtBP 1和CtBP 2的类似四聚体的组装，并且CtBP 2四聚体不稳定突变体的致癌活性是有缺陷的。为了帮助基于结构的设计工作的化合物，破坏CtBP四聚体，它是必不可少的，以了解NAD（H）如何触发四聚体组装。在这里，我们研究了NAD（H）中负责触发四聚体形成的部分。使用多角度光散射（MALS），我们表明，ADP能够促进CtBP 1和CtBP 2的四聚体形成，而AMP促进CtBP 1的四聚体组装，但不是CtBP 2。缺乏腺苷磷酸的其他NAD（H）部分，包括腺苷和掺入烟酰胺的那些，都不能促进四聚体组装。我们的CtBP 1与AMP的晶体结构揭示了磷酸腺苷在四聚体界面中的参与，精确定位了其在NAD（H）连接组装中的中心作用。CtBP 1和CtBP 2具有重叠但独特的作用，这表明对其独特结构特性的详细了解可能在设计旁系同源物特异性抑制剂中具有实用性。我们通过一系列13个位点的定点突变体研究了对AMP的不同反应。这些突变揭示了铰链段的核心作用，我们称之为120 s铰链，其连接底物与辅酶结合结构域并影响核苷酸结合和四聚体组装。我们的研究结果提供了深入了解合适的口袋，以探索基于结构的药物设计，以干扰CtBP在癌症中的共转录活性。
C-terminal binding proteins (CtBPs) are cotranscriptional factors that play key roles in cell fate. We have previously shown that NAD(H) promotes the assembly of similar tetramers from either human CtBP1 and CtBP2 and that CtBP2 tetramer destabilizing mutants are defective for oncogenic activity. To assist structure-based design efforts for compounds that disrupt CtBP tetramerization, it is essential to understand how NAD(H) triggers tetramer assembly. Here, we investigate the moieties within NAD(H) that are responsible for triggering tetramer formation. Using multiangle light scattering (MALS), we show that ADP is able to promote tetramer formation of both CtBP1 and CtBP2, whereas AMP promotes tetramer assembly of CtBP1, but not CtBP2. Other NAD(H) moieties that lack the adenosine phosphate, including adenosine and those incorporating nicotinamide, all fail to promote tetramer assembly. Our crystal structures of CtBP1 with AMP reveal participation of the adenosine phosphate in the tetrameric interface, pinpointing its central role in NAD(H)-linked assembly. CtBP1 and CtBP2 have overlapping but unique roles, suggesting that a detailed understanding of their unique structural properties might have utility in the design of paralog-specific inhibitors. We investigated the different responses to AMP through a series of site-directed mutants at 13 positions. These mutations reveal a central role for a hinge segment, which we term the 120s hinge that connects the substrate with coenzyme-binding domains and influences nucleotide binding and tetramer assembly. Our results provide insight into suitable pockets to explore in structure-based drug design to interfere with cotranscriptional activity of CtBP in cancer.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
6.2
作者：
Chawla, Ayesha T.;Chougoni, Kranthi Kumar;Grossman, Steven R.
通讯作者：
Grossman, Steven R.
DOI：
10.1073/pnas.0830998100
发表时间：
2003-04-15
影响因子：
11.1
作者：
Grooteclaes, M;Deveraux, Q;Frisch, SM
通讯作者：
Frisch, SM
影响因子：
4.8
作者：
Bellesis, Andrew G.;Jecrois, Anne M.;Royer, William E., Jr.
通讯作者：
Royer, William E., Jr.
影响因子：
--
作者：
Cubillos-Rojas M;Schneider T;Bartrons R;Ventura F;Rosa JL
通讯作者：
Rosa JL