Probing the multiple roles of the PCSK9 active site using chemical biology

利用化学生物学探讨 PCSK9 活性位点的多重作用

• 批准号: 9103195
• 负责人: John S Chorba
• 金额: $ 18.13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9103195
• 关键词: Academic Medical Centers; Active Sites; Address; Atherosclerosis; Award; Binding; Biochemical; Biochemistry; Biological; Biology; Blood Circulation; Blood Vessels; Bypass; California; Cardiology; Catalytic Domain; Cell Culture Techniques; Cell surface; Cells; Chemicals; Cholesterol; Cholesterol Homeostasis; Clinical; Cysteine; Data; Development; Disease; Dominant-Negative Mutation; Drug Targeting; Environment; Equipment; Event; Face; General Hospitals; Genetic; Goals; Grant; Health; Heart Diseases; Heel; Hepatocyte; Human; Human Genetics; In Vitro; Institution; Investigation; K-Series Research Career Programs; Kinetics; LDL Cholesterol Lipoproteins; Life Cycle Stages; Lipids; Low Density Lipoprotein Receptor; Low-Density Lipoproteins; Manuscripts; Maps; Mass Spectrum Analysis; Measurement; Mediating; Medicine; Mentors; Mentorship; Methodology; Michigan; Molecular; Organic Chemistry; Peptide Hydrolases; Pharmaceutical Preparations; Physicians; Physiology; Play; Preparation; Process; Proprotein Convertases; Protein Secretion; Proteolysis; Publishing; Reaction; Reading; Reporting; Research; Research Personnel; Research Proposals; Role; Route; San Francisco; Scientist; Serum; Signal Transduction; Site; Source; Specificity; Stroke; Substrate Specificity; Subtilisins; System; Testing; Therapeutic antibodies; Training; Translating; United States; Universities; Work; base; career; design; extracellular; hypercholesterolemia; in vitro Assay; inhibitor/antagonist; insight; lipid metabolism; loss of function; mutant; novel; novel therapeutics; prevent; professor; protein phosphatase inhibitor-2; receptor; research facility; research study; skills; skills training; small molecule; sortilin; sound; structural biology; symposium; tool; trafficking

 DESCRIPTION (provided by applicant): The candidate for this Mentored Clinical Scientist Research Career Development Award is John Chorba, MD, an Assistant Adjunct Professor (Medicine/Cardiology) at the San Francisco General Hospital and University of California, San Francisco. The primary goal of this award is for the candidate to obtain the training and skills necessary to become a successful independent physician-scientist at an academic medical center. Dr. Chorba's research objective is to use a chemical biology approach in investigating the role of the proprotein convertase subtilisin-kexin type 9 (PCSK9) active site in modulating PCSK9 processing. His long- term career goal is to use chemical tools to understand the molecular mechanisms underlying hypercholesterolemia, lipid metabolism, and the development of atherosclerosis for the purpose of developing novel therapies for heart disease. The training plan involves primary mentorship under Prof. Kevan Shokat, a world-class investigator and expert in developing chemical tools to probe biological questions. Additional guidance will be provided by an impressive group of experts who are leaders in protease biochemistry (Prof. Charles Craik, UCSF), vascular and lipid biology (Profs. Peter Ganz and John Kane, UCSF), cellular trafficking (Prof. David Ginsburg, U. of Michigan), and human genetics (Prof. Helen Hobbs, UT Southwestern). The research facilities and equipment at the institution, UCSF, are outstanding. The training plan also includes focused tutorials and didactic coursework in chemical biology, organic chemistry, structural biology, and lipid biology, as well as participatio and presentation at conferences and mentored guidance with manuscript and grant preparation. As PCSK9 acts to degrade the low-density lipoprotein (LDL) receptor, the major source of clearance of pro-atherogenic LDL cholesterol from the bloodstream, this project has direct implications for the understanding and treatment of atherosclerosis. Specifically, Aim 1 investigates the role of the PCSK9 active site in the generality and specificity of PCSK9 proteolysis, using parallel in vitro and cell culture approaches. Aim 2 investigates the impact of the active site on PCSK9 secretion and LDL-R expression, using an experimental system that bypasses the need for proteolysis in PCSK9 processing. In aim 3, the application proposes to develop chemical probes to dissect PCSK9 proteolysis from secretion and determine the structural requirements of each of these functions, and ultimately serve as starting points for novel therapeutics. Dr. Chorba's Career Development Award proposal comprises a promising candidate, outstanding training environment, rigorous training plan, and an exciting research proposal utilizing new methodologies, all with a direct relevance to human health and disease. At the completion of this Award, Dr. Chorba will have the skills necessary to succeed as an independent investigator.

 描述（由申请人提供）：本次临床科学家研究职业发展奖的候选人是约翰·乔巴（John Chorba）医学博士，他是旧金山总医院和加州大学旧金山分校的助理兼职教授（医学/心脏病学）。该奖项的目的是让候选人获得成为学术医学中心成功的独立医师科学家所需的培训和技能。 Chorba 博士的研究目标是使用化学生物学方法来研究。前蛋白转化酶枯草杆菌蛋白酶 9 (PCSK9) 活性位点在调节 PCSK9 加工中的作用他的长期职业目标是利用化学工具来了解高胆固醇血症、脂质代谢和动脉粥样硬化发展的分子机制。开发心脏病新疗法的培训计划包括凯文·肖卡特教授的主要指导，他是一位世界级的研究人员和开发化学工具来探索生物学问题的专家。令人印象深刻的专家团队，他们是蛋白酶生物化学（UCSF 教授 Charles Craik 教授）、血管和脂质生物学（UCSF 教授 Peter Ganz 和 John Kane 教授）、细胞运输（密歇根大学 David Ginsburg 教授）和人类遗传学（UT 西南大学的 Helen Hobbs 教授）的研究设施和设备非常出色。培训计划还包括化学生物学、有机学方面的重点教程和教学课程。 PCSK9 的作用是降解低密度脂蛋白 (LDL) 受体，而低密度脂蛋白 (LDL) 受体是清除促动脉粥样硬化的主要来源。血液中的 LDL 胆固醇，该项目对动脉粥样硬化的理解和治疗具有直接影响，具体而言，目标 1 研究 PCSK9 活性位点在 PCSK9 的一般性和特异性中的作用。目标 2 使用平行的体外和细胞培养方法研究活性位点对 PCSK9 分泌和 LDL-R 表达的影响，使用绕过 PCSK9 加工中蛋白水解需求的实验系统。开发化学探针来剖析 PCSK9 分泌蛋白水解作用并确定每个功能的结构要求，并最终作为新疗法的起点。发展奖提案包括有前途的候选人、出色的培训环境、严格的培训计划以及利用新方法的令人兴奋的研究提案，所有这些都与人类健康和疾病直接相关。完成该奖项后，Chorba 博士将具备相关技能。成为一名成功的独立调查员所必需的。