Paraoxonase-2 S311C Polymorphism Alters Glycosylation and Lactonase Activity

描述（由申请人提供）：此申请是为David Stoltz博士在气道细胞生物学领域的监督研究职业发展经验提供支持。Stoltz博士完成了医学博士/博士学位。目前在爱荷华大学肺病/重症监护奖学金。他的培训计划包括进一步的教学研究和细胞分子生物学和人类遗传学的实验室培训。这些经历将促进Stoltz博士成为一名独立的研究人员和杰出的内科科学家。Drs。Joseph Zabner作为主要导师，Michael Welsh作为联合导师，将负责确保职业发展计划的成功。由与拟议项目有关领域的专家组成的咨询委员会将提供持续的批判性审查以及科学和专业指导。拟开展的研究将重点关注对氧磷酶-2 （PON2）在气道上皮细胞中调控铜绿假单胞菌群体感应，这是细菌毒力和生物膜形成的重要决定因素。我们最近的研究表明，PON2使铜绿假单胞菌群体感应分子3OC12-HSL失活，并且，具有临床意义的是，PON2的常见多态性（Ser311Cys）会损害这种反应。我的初步数据表明，不同的糖基化发生在PON2变体之间。我假设PON2 Ser311 Cys多态性由于糖基化改变和/或PON2运输/定位缺陷导致内酯酶活性受损。验证这一假设的具体目的包括：1)调查哪些PON2糖基化位点发生核心糖基化和修饰，2)评估PON2变体中糖基化模式的改变是否代表PON2运输/定位的改变，以及3)确定糖基化对PON2内酯酶活性是否重要。导师和爱荷华大学都高度致力于Stoltz博士的学术成功和发展成为一名独立的研究人员。作为这一承诺的标志，他将于2007年7月加入肺/重症监护室。Stoltz博士的研究计划与公共卫生密切相关，因为铜绿假单胞菌是医院获得性感染和囊性纤维化感染和死亡的常见原因，囊性纤维化是美国最常见的遗传性致命疾病。该项目的研究结果有望为铜绿假单胞菌感染提供新的治疗选择。

DESCRIPTION (provided by applicant): This application is to provide support for a supervised research career development experience for Dr. David Stoltz in the field of airway cell biology. Dr. Stoltz has completed a combined M.D./Ph.D. program and is currently in the Pulmonary/Critical Care fellowship at the University of Iowa. His training plans include further didactic studies and laboratory training in cell and molecular biology and human genetics. These experiences will foster the development of Dr. Stoltz into an independent researcher and outstanding physician-scientist. Drs. Joseph Zabner, as primary mentor, and Michael Welsh, as co-mentor, will be responsible for ensuring the success of the career development plan. An advisory committee composed of experts in fields related to the proposed project will provide ongoing critical review and scientific as well as professional guidance. The proposed research will focus on regulation of Pseudomonas aeruginosa quorum sensing, an important determinant of bacterial virulence and biofilm formation, by paraoxonase-2 (PON2) in airway epithelial cells. We have recently shown that PON2 inactivates the P. aeruginosa quorum-sensing molecule, 3OC12-HSL and, of clinical interest, a common polymorphism in PON2 (Ser311Cys) impairs this response. My preliminary data suggest that differential glycosylation occurs between the PON2 variants. I hypothesize that the PON2 Ser311 Cys polymorphism results in impaired lactonase activity due to altered glycosylation and/or a PON2 trafficking/localization defect. The Specific Aims designed to test this hypothesis include: 1) Investigating which PON2 glycosylation sites undergo core glycosylation and modification, 2) Evaluating if the altered glycosylation pattern in the PON2 variants represents altered PON2 trafficking/localization, and 3) Determining if glycosylation is important for PON2 lactonase activity. Both the mentors and the University of Iowa are highly committed to Dr. Stoltz's academic success and development into an independent researcher. As a sign of this commitment, he will join the faculty of the Division of Pulmonary/Critical Care in July 2007. Dr Stoltz's research plan has great relevance to public health as P. aeruginosa is a common cause of infection and death in hospital acquired infections and cystic fibrosis, the most common inherited, lethal disorder in the United States. Findings from the proposed project will hopefully uncover new therapeutic options for P. aeruginosa infections.