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（由适用提供）：该指导的临床科学家研究职业发展奖的候选人是旧金山综合医院和加利福尼亚大学旧金山大学助理兼职教授（医学/心脏病学）医学博士John Chorba。该奖项的主要目标是候选人获得在学术医学中心成为成功的独立医师科学家所必需的培训和技能。 Chorba博士的研究目标是使用化学生物学方法来研究Propotein转化酶枯草蛋白 - 枯萎病型9型（PCSK9）活性位点在调节PCSK9处理中的作用。他的长期职业目标是使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的发展的分子机制，目的是开发新的心脏病疗法。该培训计划涉及世界一流的研究者凯文·索卡特（Kevan Shokat）教授的主要心态，该研究人员兼开发化学工具以探测生物学问题的专家。令人印象深刻的专家将提供其他指导，他们是蛋白酶生物化学领域（Charles Craik教授，UCSF教授），血管和脂质生物学（Peter。PeterGanz和John Kane，UCSF教授，UCSF），Cellular Fracticking，Michigan的David Ginsburg教授，以及人类的Helenentics（Hearentsick）。 UCSF机构的研究设施和设备非常出色。该培训计划还包括专注于化学生物学，有机化学，结构生物学和脂质生物学的教程和教学课程，以及在会议上的参与和演示，并通过手稿和赠款准备指导指导。由于PCSK9的作用是降解低密度脂蛋白（LDL）接收器，这是促动脉粥样硬化LDL胆固醇清除的主要来源，因此该项目对了解和治疗动脉粥样硬化具有直接的影响。具体而言，AIM 1使用平行的体外和细胞培养方法研究了PCSK9活性位点在PCSK9蛋白水解的一般性和特异性中的作用。 AIM 2使用实验系统绕过PCSK9加工中蛋白水解的需求，研究了活性位点对PCSK9分泌和LDL-R表达的影响。在AIM 3中，应用化学问题的应用建议从分泌中剖析PCSK9蛋白水解并确定每个功能的结构要求，并最终作为新治疗的起点。 Chorba博士的职业发展奖提案包括承诺候选人，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建议，所有方法都与人类健康和疾病直接相关。该奖项结束后，Chorba博士将拥有成功作为独立调查员的必要技能。