Structure and Regulation of hsp90 Chaperones

hsp90 分子伴侣的结构和调控

基本信息

批准号：
8851526
负责人：
DANIEL T GEWIRTH
金额：
$ 32.91万
依托单位：
HAUPTMAN-WOODWARD MEDICAL RESEARCH INST
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-05-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8851526
关键词：
ATP phosphohydrolase Affinity Bad protein Binding Biochemical Biological Biological Assay Cell secretion Cell surface Cells Cessation of life Client Complex Coupled Cyclic GMP-Dependent Protein Kinases Development Discrimination Disease Drug Design Exhibits Family member GRP94 Goals Grant Health Heat-Shock Proteins 90 Lectin Length Ligand Binding Ligands Malignant Neoplasms Maps Mediator of activation protein Molecular Chaperones Molecular Conformation N-terminal Neoplastic Cell Transformation Neurofibrillary Tangles Nucleotides Oncogenic Play Positioning Attribute Property Proteins Regulation Research Role Shapes Signal Transduction Staging Structure System Testing Therapeutic Time Treatment Efficacy Weather Work X-Ray Crystallography base biophysical analysis cancer cell cancer therapy cell killing in vivo inhibitor/antagonist insight neoplastic cell next generation novel osteosarcoma paralogous gene protein protein interaction response small molecule steroid hormone receptor success time use transcription factor

项目摘要

DESCRIPTION (provided by applicant): The hsp90 chaperones participate in the late stage maturation of proteins involved in diverse cellular activities ranging from signaling to bacterial recognition. Cytoplasmic Hsp90 guides the maturation of steroid hormone receptors, proto-oncogenic kinases, G-proteins, and other key mediators of neoplastic transformation. Inhibitors of Hsp90 that disrupt this maturation display potent anti-cancer activity. GRP94, the ER hsp90 paralog, chaperones proteins destined for transport to the cell surface and secretion, plays a key role in the export of toxic malfolded proteins from the ER, and when targeted leads to the death of tumor cells. The activity of hsp90 chaperones is regulated by the binding of ATP and other ligands to the N-terminal domain, and the response of this domain is central to the hsp90 chaperone cycle. Yet despite their high degree of sequence and structural homology, the cytoplasmic and ER hsp90 paralogs exhibit fundamental differences in their response to regulatory ligands. Ligand dependent conformational changes in the N-terminal domain lid of cytoplasmic Hsp90 have been difficult to demonstrate, and a plausible trigger for any changes has not been identified. In GRP94, on the other hand, a conserved insertion in the lid primes this region to undergo conformational rearrangements that differ dramatically depending on the identity of the incoming ligand. These rearrangements alter the quaternary structure of the chaperone. The research in this proposal will identify the ligand dependent conformational states available to GRP94 using X-ray crystallography to visualize the structures of ligand-driven GRP94 complexes. In vivo assays will be used to assess the biological importance of these structural rearrangements. The sensitive response of the GRP94 lid to incoming ligands allows GRP94 to bind inhibitors that cannot be accommodated in cytoplasmic Hsp90 and are therefore selective for GRP94. Because selective and non-selective ligands are closely related, we will use X-ray crystallography to visualize the both classes of ligands bound to GRP94 and Hsp90 and infer their mechanism of selectivity. Because each of the hsp90 paralogs is responsible for chaperoning distinct sets of client proteins, specific targeting of one hsp90 paralog with selective inhibitors may result in higher efficacy and therapeutic control. Finally, new developments offer the prospect of characterizing novel client and co-chaperone interactions with both Hsp90 and GRP94: minimal client protein loading systems open the way to isolating client-Hsp90 complexes for biochemical and biophysical study, and the recent identification of the first GRP94 accessory factor, os9, has, for the first time, opened the door to characterizing how protein-protein interactions occur in the ER paralog.

描述（由申请人提供）：HSP90伴侣参与参与从信号到细菌识别的各种细胞活性的蛋白质的晚期成熟。细胞质HSP90指导类固醇受体，原始性激酶，G蛋白和其他肿瘤转化的其他关键介体的成熟。破坏这种成熟的HSP90的抑制剂表现出有效的抗癌活性。 GRP94，ER HSP90旁系同源物，伴侣蛋白的蛋白质注定要运输到细胞表面和分泌，在从ER的有毒畸形蛋白出口出口中起着关键作用，而当靶向时会导致肿瘤细胞死亡。 HSP90伴侣的活性受ATP和其他配体与N末端结构域的结合调节，该结构域的响应对HSP90伴侣循环至关重要。然而，尽管它们的序列和结构同源性很高，但细胞质和ER HSP90旁系同源物在其对调节配体的响应中仍具有根本差异。细胞质HSP90的N末端结构域盖的配体依赖构象变化很难证明，并且尚未确定任何变化的合理触发因素。另一方面，在GRP94中，在盖子素中的保守插入该区域会经历构象排列，根据传入配体的身份，这些重排大不相同。这些重排改变了伴侣的第四纪结构。该提案中的研究将使用X射线晶体学来确定GRP94可用的配体依赖构象状态，以可视化配体驱动的GRP94复合物的结构。体内测定将用于评估这些结构重排的生物学重要性。 GRP94盖对传入配体的敏感响应允许GRP94结合无法在细胞质HSP90中适应的抑制剂，因此对于GRP94具有选择性。由于选择性和非选择性配体密切相关，因此我们将使用X射线晶体学来可视化与GRP94和HSP90结合的两个类别的配体，并推断它们的选择性机理。由于HSP90旁系同源物中的每一个都负责陪伴不同的客户蛋白集，因此具有选择性抑制剂的一个HSP90旁系同生的特异性靶向可能会导致更高的疗效和治疗性控制。最后，新的发展提供了表征与HSP90和GRP94：最小客户蛋白蛋白蛋白载荷系统的新型客户和联合伴侣相互作用的前景，可以为隔离客户和生物物理学研究的客户端HSP90复合物，以及对第一个GRP94辅助因子的最新识别，OS9的最新识别是开放的，该方法是开放的。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Characterization of the Grp94/OS-9 chaperone-lectin complex.

Grp94/OS-9 伴侣-凝集素复合物的表征。

DOI：
10.1016/j.jmb.2014.08.024
发表时间：
2014
期刊：
Journal of molecular biology
影响因子：
5.6
作者：
Seidler,PaulM;Shinsky,StephenA;Hong,Feng;Li,Zihai;Cosgrove,MichaelS;Gewirth,DanielT
通讯作者：
Gewirth,DanielT

Paralog Specific Hsp90 Inhibitors - A Brief History and a Bright Future.

DOI：
10.2174/1568026616666160413141154
发表时间：
2016
期刊：
Current topics in medicinal chemistry
影响因子：
3.4
作者：
Gewirth DT
通讯作者：
Gewirth DT

Exploring the Functional Complementation between Grp94 and Hsp90.

DOI：
10.1371/journal.pone.0166271
发表时间：
2016
期刊：
PloS one
影响因子：
3.7
作者：
Maharaj KA;Que NL;Hong F;Huck JD;Gill SK;Wu S;Li Z;Gewirth DT
通讯作者：
Gewirth DT

Different poses for ligand and chaperone in inhibitor-bound Hsp90 and GRP94: implications for paralog-specific drug design.

DOI：
10.1016/j.jmb.2009.03.071
发表时间：
2009-05-22
期刊：
JOURNAL OF MOLECULAR BIOLOGY
影响因子：
5.6
作者：
Immormino, Robert M.;Metzger, Louis E.;Reardon, Patrick N.;Dollins, D. Eric;Blagg, Brian S. J.;Gewirth, Daniel T.
通讯作者：
Gewirth, Daniel T.