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

 描述（由申请人提供）：该指导临床科学家研究职业发展奖的候选人是John Chorba，医学博士，旧金山弗朗西斯科总医院和加州大学旧金山分校弗朗西斯科的助理兼职教授（医学/心脏病学）。该奖项的主要目标是让候选人获得必要的培训和技能，成为学术医疗中心成功的独立医生科学家。Chorba博士的研究目标是使用化学生物学方法研究前蛋白转化酶枯草杆菌蛋白酶-kexin 9型（PCSK 9）活性位点在调节PCSK 9加工中的作用。他的长期职业目标是利用化学工具了解高胆固醇血症、脂质代谢和动脉粥样硬化发展的分子机制，以开发心脏病的新疗法为目的。 培训计划包括在Kevan Shokat教授的指导下进行初级指导，Kevan Shokat教授是一位世界级的研究人员和开发化学工具以探索生物问题的专家。额外的指导将提供一个令人印象深刻的专家组谁是蛋白酶生物化学（教授查尔斯克雷克，加州大学旧金山分校），血管和脂质生物学（教授。彼得甘茨和约翰凯恩，加州大学旧金山分校），细胞贩运（教授大卫金斯伯格，美国。和人类遗传学（Helen Hobbs教授，UT Southwestern）。加州大学旧金山分校的研究设施和设备非常出色。培训计划还包括化学生物学，有机化学，结构生物学和脂质生物学的重点教程和教学课程，以及在会议上的参与和演示，以及指导手稿和赠款准备。 由于PCSK 9可降解低密度脂蛋白（LDL）受体，这是从血液中清除促动脉粥样硬化LDL胆固醇的主要来源，因此该项目对动脉粥样硬化的理解和治疗具有直接意义。具体而言，目的1研究PCSK 9活性位点在PCSK 9蛋白水解的一般性和特异性中的作用，使用平行的体外和细胞培养方法。目的2研究活性位点对PCSK 9分泌和LDL-R表达的影响，使用绕过PCSK 9加工中蛋白水解需要的实验系统。在目标3中，本申请提出开发化学探针以从分泌中剖析PCSK 9蛋白水解，并确定这些功能中的每一个的结构要求，并最终作为新疗法的起点。 Chorba博士的职业发展奖提案包括一个有前途的候选人，出色的培训环境，严格的培训计划，以及一个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。在这个奖项完成后，Chorba博士将拥有必要的技能，成功地作为一个独立的调查员。

 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.