Structures of PGAM5 Provide Insight into Active Site Plasticity and Multimeric Assembly.
Structures of PGAM5 Provide Insight into Active Site Plasticity and Multimeric Assembly.
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DOI：
10.1016/j.str.2017.05.020
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发表时间：
2017-07-05
期刊：
影响因子：
--
通讯作者：
Knapp S
中科院分区：
文献类型：
--
作者：
Chaikuad A;Filippakopoulos P;Marcsisin SR;Picaud S;Schröder M;Sekine S;Ichijo H;Engen JR;Takeda K;Knapp S
PGAM 5是一种线粒体膜蛋白，其作为非典型Ser/Thr磷酸酶发挥功能，并且是氧化应激反应、坏死性凋亡和自噬的调节剂。在这里，我们提出了几个晶体结构的PGAM 5，包括激活N-末端调控序列，提供了一个模型的结构可塑性，催化结构域的二聚体，并组装成一个酶活性的十二聚体形式。在结构中观察到的低聚状态得到氢交换质谱法，尺寸排阻色谱法和溶液中的分析超浓缩实验的支持。我们报告说，催化重要的N-末端WDPNWD基序作为一个结构整合器组装成一个十二聚体的PGAM 5，变构激活的磷酸酶，促进有序的催化环。此外，所观察到的活性位点的可塑性，使催化元素的基本构象重排的可视化。全面的生物物理表征提供了这种关键的线粒体磷酸酶的详细结构模型，这种磷酸酶与多种疾病的发展有关。PGAM 5催化结构域共享磷酸甘油酸三倍并形成稳定的二聚体WDPNWD基序变构激活完全活性的十二聚体形式晶体结构揭示了PGAM 5活性位点的构象可塑性Chaikuad et al.提出了PGAM 5的多种晶体结构及其在溶液中的生物物理分析，揭示了调节寡聚化及其变构激活的可塑性和结构机制。
PGAM5 is a mitochondrial membrane protein that functions as an atypical Ser/Thr phosphatase and is a regulator of oxidative stress response, necroptosis, and autophagy. Here we present several crystal structures of PGAM5 including the activating N-terminal regulatory sequences, providing a model for structural plasticity, dimerization of the catalytic domain, and the assembly into an enzymatically active dodecameric form. Oligomeric states observed in structures were supported by hydrogen exchange mass spectrometry, size-exclusion chromatography, and analytical ultracentrifugation experiments in solution. We report that the catalytically important N-terminal WDPNWD motif acts as a structural integrator assembling PGAM5 into a dodecamer, allosterically activating the phosphatase by promoting an ordering of the catalytic loop. Additionally the observed active site plasticity enabled visualization of essential conformational rearrangements of catalytic elements. The comprehensive biophysical characterization offers detailed structural models of this key mitochondrial phosphatase that has been associated with the development of diverse diseases. PGAM5 catalytic domain shares phosphoglycerate mutase fold and forms stable dimer WDPNWD motif allosterically activates the fully active dodecameric form Crystal structures reveal conformational plasticity of the PGAM5 active site Chaikuad et al. present multiple crystal structures of PGAM5 and its biophysical analysis in solution, revealing the plasticity and structural mechanisms regulating oligomerization and its allosteric activation.
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DOI：
10.1107/s090744491003982x
发表时间：
2011-04
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Evans PR
通讯作者：
Evans PR
影响因子：
4.5
作者：
Imai Y;Kanao T;Sawada T;Kobayashi Y;Moriwaki Y;Ishida Y;Takeda K;Ichijo H;Lu B;Takahashi R
通讯作者：
Takahashi R
影响因子：
3.7
作者：
Ishida Y;Sekine Y;Oguchi H;Chihara T;Miura M;Ichijo H;Takeda K
通讯作者：
Takeda K
影响因子：
44.1
作者：
通讯作者：
--
影响因子：
2.9
作者：
Chen, Yunting;Jakoncic, Jean;Nassar, Nicolas
通讯作者：
Nassar, Nicolas