Paraoxonase-2 S311C Polymorphism Alters Glycosylation and Lactonase Activity

描述（由申请人提供）：本申请旨在为大卫斯托尔兹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博士已经完成了医学博士/博士项目，目前在爱荷华州大学的肺/重症监护奖学金。他的培训计划包括在细胞和分子生物学以及人类遗传学方面的进一步教学研究和实验室培训。这些经验将促进Stoltz博士发展成为一名独立的研究人员和杰出的物理学家。约瑟夫Zabner博士，作为主要导师，和迈克尔威尔士，作为共同导师，将负责确保职业发展计划的成功。一个由拟议项目相关领域专家组成的咨询委员会将提供持续的严格审查和科学及专业指导。拟议的研究将集中在调节铜绿假单胞菌群体感应，细菌毒力和生物膜形成的重要决定因素，对氧磷酶2（PON 2）在气道上皮细胞。我们最近发现PON 2使铜绿假单胞菌群体感应分子3 OC 12-HSL失活，并且临床上感兴趣的是PON 2（Ser 311 Cys）中的常见多态性损害这种反应。我的初步数据表明，差异糖基化发生在PON 2变体之间。我推测PON 2 Ser 311 Cys多态性导致内酯酶活性受损，这是由于糖基化改变和/或PON 2运输/定位缺陷。设计用于检验该假设的具体目的包括：1）研究哪些PON 2糖基化位点经历核心糖基化和修饰，2）评价PON 2变体中改变的糖基化模式是否代表改变的PON 2运输/定位，以及3）确定糖基化是否对PON 2内酯酶活性重要。导师和爱荷华州大学都高度致力于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/重症监护科。Stoltz博士的研究计划与公共卫生有很大的相关性，因为铜绿假单胞菌是医院获得性感染和囊性纤维化中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。该项目的发现有望为铜绿假单胞菌感染发现新的治疗选择。

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.