Elucidating structural, mechanistic, and allosteric determinants of mTOR Complex 2 (mTORC2) signaling.

阐明 mTOR 复合物 2 (mTORC2) 信号传导的结构、机制和变构决定因素。

• 批准号: 10450814
• 负责人: MARTIN S TAYLOR
• 金额: $ 17.11万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10450814
• 关键词: Address; Affect; Affinity; Aging; Allosteric Regulation; Apoptosis; Autophagocytosis; Award; Binding; Biochemical; Bioinformatics; Biological Assay; Biology; Biomedical Research; Catabolic Process; Catalysis; Chemicals; Clinical; Collaborations; Color; Complex; Cryoelectron Microscopy; Data; Data Analyses; Development; Development Plans; Diabetes Mellitus; Disease; Ensure; Eukaryotic Cell; FRAP1 gene; Feedback; Fostering; Functional disorder; General Hospitals; Generations; Glucose Transporter; Goals; Growth; Growth Factor; Homeostasis; Hospital Departments; Human; Hypertension; Impairment; Institutes; Institution; Insulin; Insulin Receptor; Insulin Resistance; International; Ion Channel; Kinetics; Knowledge; Learning; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Massachusetts; Measures; Mentors; Metabolic; Metabolic syndrome; Molecular Conformation; Monomeric GTP-Binding Proteins; Neoplasm Metastasis; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Oncogenic; Pathologist; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phosphorylation; Phosphotransferases; Physicians; Play; Positioning Attribute; Principal Investigator; Protein Engineering; Proteins; Proteomics; Reaction; Reagent; Regulation; Research; Research Personnel; Resolution; Role; Science; Scientist; Series; Signal Transduction; Solid Neoplasm; Structure; Substrate Interaction; System; Techniques; Testing; Therapeutic; Time; Training; Treatment Failure; Tuberous Sclerosis; Universities; Up-Regulation; Visualization; arm; base; blood glucose regulation; cancer type; career; career development; cell growth; cell motility; chemical conjugate; crosslink; density; design; experience; follow-up; gastrointestinal; glucose transport; human disease; inhibitor; insight; insulin sensitivity; insulin signaling; interest; mTOR inhibition; nanobodies; novel; prevent; programs; protein complex; receptor sensitivity; research and development; rho; rho GTP-Binding Proteins; skills; small molecule; structural biology; success; therapy development; tool

Taylor, Martin | K08 (PA-20-203) | PROJECT SUMMARY / ABSTRACT This proposal details a five-year training plan for the development of a research program focused on elucidating structural, mechanistic, and allosteric determinants of mTOR Complex 2 (mTORC2) signaling. Phosphorylation of the signaling kinase Akt on Ser473 by mTOR Complex 2 (mTORC2) is a critical regulated intracellular step in insulin and other growth factor signaling. Ser473 phosphorylation activates Akt and is required for Akt’s downstream metabolic effects, such as glucose transporter upregulation, which are dysregulated in diabetes, and proliferation and growth, which are dysregulated in cancer. Therapeutic modulation of mTORC2 is therefore of interest in the treatment of both diabetes and cancer but is not yet possible due to similarities between mTORC2 and the better-understood effector complex mTORC1, which shares key components mTOR and mLST8. mTORC2 also activates other substrates with less-defined roles in ion channel homeostasis, apoptosis, cell motility, metastasis, and insulin receptor sensitivity, and its activity is modulated by a series of allosteric interacting proteins including the small GTPases Rho and Ras. However, despite the central role of mTORC2- Akt signaling, we have little information about how this critical reaction is catalyzed by mTORC2, how mTORC2 recognizes Akt and other substrates, or how these interactions are modulated allosterically. This project therefore seeks to develop a detailed structural and mechanistic understanding of mTORC2 recognition of its substrates, using novel enzymologic assays, protein engineering, and a combination of biochemical, chemical, proteomic, and structural biology approaches that will also develop proof-of-concept inhibitors and potentially activators of mTORC2. My proposed studies will: (i) provide detailed structural and mechanistic insight into mTOR Complex 2 kinase signaling (ii) develop tools, reagents, and techniques applicable to other systems of interest, including mTORC1 (iii) provide ample opportunities for mechanistic and translational follow-up for my transition to independence. I am a practicing gastrointestinal pathologist and physician scientist seeking K08 support for mentored research under the guidance of Dr. David Sabatini and Dr. Philip Cole. This mentored period of 80% research and career development and 20% clinical time will ensure I acquire the skills required to become a successful independent principal investigator. Drs. Sabatini and Cole are internationally recognized mentors, together training ~40 successful independent investigators. My training will occur at two world-class institutions, the Whitehead Institute for Biomedical Research, and Massachusetts General Hospital Department of Pathology. Both are rich with opportunities for young scientists to train, pursue highly impactful science, and foster long-lasting collaborations. I will also be guided by a committee of researchers that are all leaders in their fields: Dr. Joseph Davis (MIT, structural biology of large protein complexes and cryoEM data analysis) and Dr. Yi Shi (University of Pittsburgh, crosslinking mass spectrometry as applied to large protein complexes and nanobody generation). The support of this K08 award will allow me to focus on maturing my research and strengthening my career development during this critical last stage of mentored training. At the conclusion of my award period, I will be optimally positioned for achieving success as an independent physician scientist.

泰勒，马丁| K08（PA-20-203）|项目摘要 /摘要 该建议详细介绍了一个为期旨在阐明的研究计划的五年培训计划 MTOR复合物2（MTORC2）信号传导的结构，机械和变构决定器。磷酸化 MTOR复合物2（MTORC2）在Ser473上的信号传导激酶AKT的akt是关键的调控内阶跃 胰岛素和其他生长因子信号传导。 SER473磷酸化激活AKT，是Akt所必需的 下游代谢作用，例如葡萄糖转运蛋白上调，在糖尿病中失调， 以及在癌症中失调的增殖和生长。因此，MTORC2的治疗调节是 对糖尿病和癌症的治疗感兴趣，但由于之间的相似性是不可能的 MTORC2和更好理解的效应综合体MTORC1，共享密钥组件MTOR和 MLST8。 MTORC2还激活其他底物，在离子通道体内稳态，凋亡，凋亡， 细胞运动，转移和胰岛素受体敏感性，其活性由一系列变构调节 相互作用的蛋白质，包括小gtpase rho和ras。但是，尽管MTORC2- AKT信号，我们几乎没有关于MTORC2如何催化这种关键反应的信息 识别AKT和其他底物，或如何对这些相互作用进行变构调节。因此，这个项目 试图对MTORC2识别其底物的识别，详细的结构和机械理解， 使用新型的酶学测定，蛋白质工程以及生化，化学，蛋白质组学的组合 以及还将开发出概念验证抑制剂和潜在激活因子的结构生物学方法 mtorc2。 我建议的研究将： （i）对MTOR复合物2激酶信号提供详细的结构和机械洞察力 （ii）开发适用于其他感兴趣系统的工具，试剂和技术，包括MTORC1 （iii）为我向独立过渡提供了足够的机械和翻译跟进机会。 我是胃肠道病理学家和身体科学家的实践，寻求K08支持指导研究 在戴维·萨巴蒂尼（David Sabatini）博士和菲利普·科尔（Philip Cole）博士的指导下。这是80％的研究和职业生涯的时期 开发和20％的临床时间将确保我获得成功独立的技能 首席研究员。博士。萨巴蒂尼（Sabatini）和科尔（Cole）是国际认可的导师，共同培训〜40 成功的独立研究人员。我的培训将在两个世界一流的机构（怀特海）进行 生物医学研究所和马萨诸塞州综合医院病理学系。两者都是富有的 有机会为年轻科学家训练，追求高度影响力的科学，并培养长期 协作。我还将受到研究人员委员会的指导，这些研究人员都是他们领域的领导者：约瑟夫博士 戴维斯（麻省理工学院，大型蛋白质复合物的结构生物学和冷冻数据分析）和Yi Shi博士（大学 匹兹堡的，将质谱的交联质谱法应用于大型蛋白质复合物和纳米病的产生）。 这个K08奖的支持将使我能够专注于成熟并加强职业生涯 在修改培训的最后阶段的关键最后阶段的发展。在颁奖期结束时，我将 作为独立物理科学家成功的最佳位置。