Nucleotide Binding and Active Site Gate Dynamics for the Hsp90 Chaperone ATPase Domain from Benchtop and High Field F-19 NMR Spectroscopy

Rashid Suad; Lee Brian L.; Wajda Benjamin; Spyracopoulos Leo

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Nucleotide Binding and Active Site Gate Dynamics for the Hsp90 Chaperone ATPase Domain from Benchtop and High Field F-19 NMR Spectroscopy

机译：HSP90伴侣ATP酶从台式和高场F-19 NMR光谱学的HSP90伴侣ATP酶域的核苷酸结合和活性位点动态

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Protein turnover in cells is regulated by the ATP dependent activity of the Hsp90 chaperone. In concert with accessory proteins, ATP hydrolysis drives the obligate Hsp90 dimer through a cycle between open and closed states that is critical for assisting the folding and stability of hundreds of proteins. Cycling is initiated by ATP binding to the ATPase domain, with the chaperone and the active site gates in the dimer in open states. The chaperone then adopts a short-lived, ATP bound closed state with a closed active site gate. The structural and dynamic changes induced in the ATPase domain and active site gate upon nucleotide binding, and their impact on dimer closing are not well understood. We site-specifically (19)Flabeled the ATPase domain at the active site gate to enable benchtop and high field F-19 NMR spectroscopic studies. Combined with MD simulations, this allowed accurate characterization of pico- to nanosecond time scale motions of the active site gate, as well as slower micro- to millisecond time scale processes resulting from nucleotide binding. ATP binding induces increased flexibility at one of the hinges of the active site gate, a necessary prelude to release of the second hinge and eventual gate closure in the intact chaperone.

机译：细胞中的蛋白质转换由HSP90伴侣酮的ATP依赖性活性调节。在辅助蛋白的音乐会中，ATP水解通过开放和封闭状态之间的循环驱动了迫使的HSP90二聚体，这对于辅助数百种蛋白质的折叠和稳定性至关重要。通过ATP与ATPase结构域的ATP结合引发循环，伴侣酮和在开放状态下的二聚体中的活性位点栅极。然后伴侣伴侣采用短寿命的ATP束缚闭合状态，带有封闭的活动场地门。在核苷酸结合时，在ATP酶结构域和活性位点栅中诱导的结构和动态变化及其对二聚体闭合的影响尚不清楚。我们的网站 - 特别是（19）将ATPase域镶嵌在活动场地门处，以使得能够高压和高场F-19 NMR光谱研究。结合MD仿真，这允许精确表征了活性位点栅极的微微纳入纳秒时间尺度运动，以及由核苷酸结合引起的微量达毫秒的时间尺度过程。 ATP绑定在有源位点栅极的一个铰链中引起增加的灵活性，该铰链的铰链是在完整的伴侣中释放第二铰链和最终栅极封闭的必要前列。

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《The journal of physical chemistry, B. Condensed matter, materials, surfaces, interfaces & biophysical》 |2020年第15期|共10页
作者
Rashid Suad; Lee Brian L.; Wajda Benjamin; Spyracopoulos Leo;
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Univ Alberta Dept Biochem Edmonton AB T6G 2H7 Canada;

Univ Alberta Dept Biochem Edmonton AB T6G 2H7 Canada;

Univ Alberta Dept Biochem Edmonton AB T6G 2H7 Canada;

Univ Alberta Dept Biochem Edmonton AB T6G 2H7 Canada;

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正文语种 eng
中图分类物理化学（理论化学）、化学物理学;
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1. Nucleotide Binding and Active Site Gate Dynamics for the Hsp90 Chaperone ATPase Domain from Benchtop and High Field F-19 NMR Spectroscopy [J] . Rashid Suad, Lee Brian L., Wajda Benjamin, The journal of physical chemistry, B. Condensed matter, materials, surfaces, interfaces & biophysical . 2020,第15期

机译：HSP90伴侣ATP酶从台式和高场F-19 NMR光谱学的HSP90伴侣ATP酶域的核苷酸结合和活性位点动态
2. Conformational Changes of Nucleotide Binding Sites Following Sequential Addition of ADP to Nucleotide-depleted Fi-ATPase of Escherichia coli Investigated with ~(31)P NMR Spectroscopy [J] . Joon Hyung Sohn, Namkung Jun, Byung-II Yeh Bulletin of the Korean Chemical Society . 2011,第3期

机译：〜（31）P NMR光谱法研究将ADP顺序添加至大肠杆菌的核苷酸耗尽型Fi-ATPase后核苷酸结合位点的构象变化
3. Conformational Changes of Nucleotide Binding Sites Following Sequential Addition of ADP to Nucleotide-depleted F₁-ATPase of Escherichia coli Investigated with 31P NMR Spectroscopy [J] . Byung Il Yeh, Hyun Won Kim, Jong Whan Choi, Bulletin of the Korean Chemical Society . 2011,第3期

机译：用 31 P NMR光谱法研究将ADP顺序添加至大肠杆菌的核苷酸耗尽型F _{1 -ATPase后核苷酸结合位点的构象变化}

4. Low-Field RheoNMR: Newly Developed Combination of Rheology and Time Domain (TD)-NMR to Correlate Mechanical Properties with Molecular Dynamics in Polymer Melts [C] . Volker R?ntzsch, Mürüvvet Begüm ?zen, Karl-Friedrich Ratzsch, International Conference of the Polymer Processing Society . 2017

机译：低场rheonmr：新开发的流变学和时域（Td）-nmr与聚合物熔体中分子动力学相关的机械性能

5. The Myosin-Binding UCS Domain but not the Hsp90-Binding TPR Domain of the UNC-45 Chaperone is Essential for Myosin Accumulation and Assembly in Caenorhabditis elegans [D] . Ni, Weiming. 2011

机译：肌苷结合的UCS结构域但不是UNC-45伴侣的HSP90结合TPR结构域对于Caenorhabditis elegans中的肌球蛋白积累和组装是必不可少的

6. Atomistic simulations and network-based modeling of the Hsp90-Cdc37 chaperone binding with Cdk4 client protein: A mechanism of chaperoning kinase clients by exploiting weak spots of intrinsically dynamic kinase domains [O] . Josh Czemeres, Kurt Buse, Gennady M. Verkhivker 2011

机译：Hsp90-Cdc37伴侣蛋白与Cdk4客户蛋白结合的原子模拟和基于网络的建模：通过利用内在动态激酶结构域的弱点来伴侣蛋白激酶客户的机制

7. Atomistic simulations and network-based modeling of the Hsp90-Cdc37 chaperone binding with Cdk4 client protein: A mechanism of chaperoning kinase clients by exploiting weak spots of intrinsically dynamic kinase domains. [O] . Josh Czemeres, Kurt Buse, Gennady M Verkhivker 2017

机译：Hsp90-Cdc37伴侣与Cdk4客户蛋白结合的原子模拟和基于网络的建模：通过利用内在动态激酶结构域的弱点来陪伴激酶客户的机制。

8. Structural and Functional Studies of Fungal Cellulose-Binding Domains by NMR211 Spectroscopy [R] . Mattinen, M. L. 1998

机译：通过NmR211光谱学对真菌纤维素结合域的结构和功能研究

1. 新异吲哚衍生物，含有它们的组合物，它们的制备与其药物用途，特别作为Hsp90伴侣蛋白活性抑制剂的药物用途 [P] . 中国专利： CN101287733A . 2008-10-15

2. method for inhibiting the activity of a separate site of an aptamer binding site in a ligand, methods of increasing the antagonist, receptor, ligand-binding aptamer, antagonist, ligand of a receptor-binding aptamer a vegf aptamer, and a vegfr aptamer ligand, method to increase an antagonist property of an aptamer that targets a protein that interacts with a second protein, method to identify an aptamer conjugate, methods to release a biologically molecule active, nucleic acid, an aptamer, and an anti-vegf aptamer to an eye, compound, composition to release a biologically active molecule to an eye [P] . 外国专利： BRPI0509911A . 2007-09-18

机译：抑制配体中适体结合位点的单独位点的活性的方法，增加拮抗剂，受体，配体结合适体，拮抗剂，受体结合适体的配体，vegf适体和vegfr适体配体的方法，增加靶向与第二种蛋白质相互作用的蛋白质的适体的拮抗特性的方法，鉴定适体缀合物的方法，将具有生物活性的分子，核酸，适体和抗vegf适体释放到眼睛的方法，化合物，成分将生物活性分子释放到眼睛

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机译：利用特定氨基酸标记的蛋白质和一维核磁共振技术筛选目标蛋白质活性位点的化合物结合方法

4. METHOD FOR SCREENING COMPOUND BINDING TO THE ACTIVE SITE OF TARGET PROTEIN BY USING PROTEIN LABELED ON SPECIFIC AMINO ACIDS AND 2D NMR TECHNIQUE [P] . 外国专利： KR100456054B1 . 2004-11-10

机译：利用特定氨基酸标记的蛋白质和二维核磁共振技术筛选目标蛋白质活性位点的化合物结合方法

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Nucleotide Binding and Active Site Gate Dynamics for the Hsp90 Chaperone ATPase Domain from Benchtop and High Field F-19 NMR Spectroscopy

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