MOLECULAR CONTROL OF CALCIUM INFLUX AT THE ER-PLASMA MEMBRANE JUNCTIONS

ER-血浆膜连接处钙流入的分子控制

• 批准号: 10386835
• 负责人: Yubin Zhou
• 金额: $ 29.18万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386835
• 关键词: Adopted; Animal Model; Biochemical; Biological Process; Biophysics; Calcium; Calcium Channel; Calcium Signaling; Cardiovascular Diseases; Cell membrane; Cells; Cellular Metabolic Process; Clinical; Clinical Medicine; Communication; Complex; Coupling; Data; Development; Disease; Drug Targeting; EF Hand Motifs; Exposure to; Functional disorder; Gene Expression; Generations; Genetic; Goals; Homeostasis; Human; Immunologic Deficiency Syndromes; Kinetics; Knock-out; Knockout Mice; Knowledge; Label; Laboratories; Link; Location; Lymphocyte; Lymphocyte Activation; Mammals; Maps; Membrane; Membrane Fusion; Mission; Modeling; Molecular; Muscle; Muscle Contraction; Neoplasm Metastasis; Pathologic; Pathway interactions; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositol Phosphates; Phosphatidylinositols; Physiological; Physiological Processes; Pilot Projects; Protein Engineering; Protein Family; Proteins; Proteomics; Public Health; Research; Route; STIM1 gene; Severe Combined Immunodeficiency; Shapes; Side; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Stimulus; Structure-Activity Relationship; System; T-Cell Activation; Techniques; Testing; Therapeutic; Therapeutic Intervention; Transgenic Mice; Translating; Tubular Aggregate Myopathies; United States National Institutes of Health; base; clinically relevant; disability; druggable target; extracellular; gain of function mutation; human disease; innovation; insight; mouse model; novel; optogenetics; overexpression; public health relevance; therapeutic target; therapeutically effective; tool; trafficking

Project Summary / Abstract. Store-operated calcium entry (SOCE) constitutes a major calcium entry pathway in mammals to control lymphocyte activation, muscle contraction, gene expression and cell metabolism. The calcium release- activated calcium (CRAC) channel composed of ORAI-STIM represents a prototypical example of SOCE in lymphocytes. The clinical relevance of SOCE is exemplified by two human diseases, the severe combined immunodeficiency (SCID) and tubular aggregate myopathy (TAM), which are caused by loss- or gain-of-function mutations in ORAI1 and STIM1, respectively. Augmented SOCE is also implicated in cardiovascular disorders and cancer metastasis. Therefore, CRAC channel has been pursued as an attractive drug target for therapeutic intervention. Tremendous efforts have been directed to establish ORAI-STIM as the minimal two-component system to couple ER calcium store depletion with calcium influx across the plasma membrane. The regulatory machinery dedicated to the ORAI-STIM signaling, nonetheless, still remains incompletely defined. In this proposal, the PI aims to bridge this critical knowledge gap by unveiling the functions of two novel SOCE modulators, which reside at distinct subcellular locations to act on different steps of ORAI-STIM signaling: the initial activation of STIM within the ER lumen and the later stabilization of ORAI-STIM complexes at ER-PM membrane contact sites (MCS), where the close appositions of two membranes are separated by a gap distance of 10-30 nm. In Aim 1, based on preliminary findings from proteomic profiling of potential STIM1 interactors within the ER lumen, the PI will define how a previously-unrecognized multiple EF-hand protein cooperates with the luminal domain of STIM1 (EFSAM) to shape the activation and deactivation kinetics of SOCE. The PI will employ a new “ER-to-PM” trafficking strategy to expose the luminal domain toward the extracellular side, thereby overcoming a major impediment to studies on the liminal sides of ER-resident signaling proteins. In Aim 2, capitalizing on the discovery of a TMEM family protein as a regulator of calcium influx at ER-PM MCS, the PI will define how this modulator responds to physiological stimuli to remodel the assembly of ER-PM junctions and PIP homeostasis to sustain SOCE. The generation of innovative optogenetic tools and a transgenic mouse model to dissect calcium signaling and protein-PIP interactions will further accelerate our structure-function relationship studies on these novel regulators. Overall, the new mechanistic insights gained through the proposed study will lead to advances in the constantly-revitalized field of calcium signaling, and in parallel, spawn the vibrant field of membrane contact sites. In the long run, discoveries made in the study can be translated into the development of effective therapeutics targeting aberrant calcium and phosphoinositide signaling.

项目摘要/摘要。 钙库操纵的钙进入（SOCE）是哺乳动物控制钙进入的主要途径 淋巴细胞激活、肌肉收缩、基因表达和细胞代谢。 由 ORAI-STIM 组成的活化钙 (CRAC) 通道代表了 SOCE 的典型例子 SOCE 的临床相关性以两种人类疾病为例，即严重的联合疾病。 免疫缺陷（SCID）和管状聚集性肌病（TAM），由功能丧失或获得引起 ORAI1 和 STIM1 的突变也与心血管疾病有关。 因此，CRAC通道已被视为有吸引力的治疗药物靶点。 为了将 ORAI-STIM 建立为最小的两部分，我们付出了巨大的努力。 将内质网钙储备耗尽与钙跨质膜流入耦合的系统。 然而，专用于 ORAI-STIM 信号传导的机制仍然不完全定义。 在这项提案中，PI 旨在通过揭示两本小说的功能来弥合这一关键知识差距 SOCE 调节剂，位于不同的亚细胞位置，作用于 ORAI-STIM 的不同步骤 信号传导：ER 腔内 STIM 的初始激活和 ORAI-STIM 复合物的后期稳定 在 ER-PM 膜接触位点 (MCS)，其中两个膜的紧密并置被一个 间隙距离为 10-30 nm 在目标 1 中，基于潜在 STIM1 蛋白质组学分析的初步发现。 ER 腔内的相互作用因子，PI 将定义以前未被识别的多重 EF 手蛋白 与 STIM1 (EFSAM) 的管腔结构域配合，塑造 STIM1 的激活和失活动力学 SOCE 将采用新的“ER-to-PM”贩运策略，将 luminal 域暴露给 细胞外侧，从而克服了 ER 驻留阈侧研究的主要障碍 在目标 2 中，利用 TMEM 家族蛋白作为钙调节剂的发现。 当 ER-PM MCS 流入时，PI 将定义该调节器如何响应生理刺激以重塑 ER-PM 连接和 PIP 稳态的组装以维持 SOCE 创新光遗传学的产生。 剖析钙信号传导和蛋白质-PIP 相互作用的工具和转基因小鼠模型将进一步 加速我们对这些新型调节剂的结构与功能关系的研究。 总体而言，通过拟议的研究获得的新的机制见解将导致在 钙信号传导场不断焕发活力，同时产生充满活力的膜接触场 从长远来看，研究中的发现可以转化为有效性的发展。 针对异常钙和磷酸肌醇信号传导的治疗方法。

项目成果

An Engineered Split-TET2 Enzyme for Chemical-inducible DNA Hydroxymethylation and Epigenetic Remodeling.

一种用于化学诱导 DNA 羟甲基化和表观遗传重塑的工程化 Split-TET2 酶。

• DOI: 10.3791/56858
• 发表时间: 2017-12-18
• 期刊: Journal of visualized experiments : JoVE
• 作者: Minjung Lee;Yubin Zhou;Yun Huang
• 通讯作者: Yun Huang

Optogenetic engineering to probe the molecular choreography of STIM1-mediated cell signaling.

光遗传学工程探索 STIM1 介导的细胞信号传导的分子编排。

• 发表时间: 2020
• 影响因子: 16.6
• 作者: Ma, Guolin;He, Lian;Liu, Shuzhong;Xie, Jiansheng;Huang, Zixian;Jing, Ji;Lee, Yi;Wang, Rui;Luo, Hesheng;Han, Weidong;Huang, Yun;Zhou, Yubin
• 通讯作者: Zhou, Yubin

Characterization of the dual functional effects of heat shock proteins (HSPs) in cancer hallmarks to aid development of HSP inhibitors.

表征热休克蛋白 (HSP) 在癌症标志中的双重功能效应，以帮助开发 HSP 抑制剂。

• 发表时间: 2020-11-23
• 影响因子: 12.3
• 作者: Zhang, Zhao;Jing, Ji;Ye, Youqiong;Chen, Zhiao;Jing, Ying;Li, Shengli;Hong, Wei;Ruan, Hang;Liu, Yaoming;Hu, Qingsong;Wang, Jun;Li, Wenbo;Lin, Chunru;Diao, Lixia;Zhou, Yubin;Han, Leng
• 通讯作者: Han, Leng

Novel intramolecular photoinduced electron transfer-based probe for the Human Ether-a-go-go-Related Gene (hERG) potassium channel.

用于人类 Ether-a-go-go 相关基因 (hERG) 钾通道的新型分子内光诱导电子转移探针。

• 发表时间: 2015-12-21
• 作者: Liu Z;Zhou Y;Du L;Li M
• 通讯作者: Li M

Inside-out Ca(2+) signalling prompted by STIM1 conformational switch.

由 STIM1 构象转换引发的由内而外的 Ca(2 ) 信号传导。

• 发表时间: 2015-07-17
• 影响因子: 16.6
• 作者: Ma, Guolin;Wei, Ming;He, Lian;Liu, Chongxu;Wu, Bo;Zhang, Shenyuan L;Jing, Ji;Liang, Xiaowen;Senes, Alessandro;Tan, Peng;Li, Siwei;Sun, Aomin;Bi, Yunchen;Zhong, Ling;Si, Hongjiang;Shen, Yuequan;Li, Minyong;Lee, Mi;Zhou, Weibin;Wang
• 通讯作者: Wang