Lysosomal Phospholipase A2 in Lung DPPC

首席调查员：费希尔。阿伦·B。 项目6：肺DPPC代谢中的溶酶体磷脂酶A2 名称eraCommons在项目从属关系中的角色 阿伦·B·费舍尔，医学博士，负责任的投资。大学。宾夕法尼亚大学 谢尔顿·I·范斯坦，博士，范斯坦大学联合研究员。宾夕法尼亚大学 耶菲姆·马内维奇博士，大学联合调查员。宾夕法尼亚大学 布莱恩·班森，博士，参与大学。删除 调查员 大江友友，博士，参与大学。宾夕法尼亚大学 调查员 职位 教授兼所长 高级研究投资公司。 高级研究投资公司。 副教授 助理教授 摘要 本项目将评估一种新的磷脂酶A2(AiPLA2)的性质和生理作用 过氧化还蛋白6的酶活性。我们从大鼠和 牛肺，已经鉴定出人、大鼠、小鼠和牛酶的全长cdna，有 产生了重组蛋白，并开发出了一组对Western有效的抗体 免疫印迹、免疫沉淀和免疫细胞化学。AiPLA2不依赖于钙离子，显示最大活性 在pH为4时，代表一种溶酶体型PLA2。在目前的赠款支助期间，我们已经表明 AiPLA2：1)在内化的肺泡DPPC的降解中起主要作用；2)提供底物 DPPC合成的酰化途径：3)活性通过与SP-A的蛋白质-蛋白质相互作用来调节； 4)定位于肺中的板层小体和溶酶体；5)被磷酸化激活；以及6)其 表达是发育调节的，并对激素有反应。特定目标1将研究以下模型 AiPLA2(基因敲除和转基因小鼠)的低表达和高表达以评价其生理作用 利用分离的细胞、分离的灌流肺和体内研究该酶在DPPC降解和合成中的作用 学习。将确定肺磷脂含量随时间(年龄)的变化。特定目标2将 评价aiPLA2活性利用的结构-功能关系

Principal Investigator: Fisher. Aron B. PROJECT #6: LYSOSOMAL PHOSPHOLIPASE A2 IN LUNG DPPC METABOLISM Name eraCommons Role on Project Affiliation Aron B. Fisher, M.D. abfifem Responsible Invest. Univ. Penn Sheldon I. Feinstein, Ph.D. feinsteins Co-Investigator Univ. Penn Yefim Manevich, Ph.D. Co-Investigator Univ. Penn Brian Bahnson, Ph.D. Participating Univ. Del Investigator Tomo Oe, Ph.D. Participating Univ. Penn Investigator Position Professor and Director Sr. Research Invest. Sr. Research Invest. Associate Professor Res. Asst. Professor ABSTRACT This project will evaluate the properties and physiological role of a novel phospholipase A2 (aiPLA2) that is one of the enzymatic activities of the protein called peroxiredoxin 6. We have isolated the protein from rat and bovine lungs, have identified the full length cDNA for the human, rat, mouse and bovine enzyme, have generated recombinant protein, and have developed a panel of antibodies that are effective for western blotting, immunoprecipitation and immunocytochemistry. aiPLA2 is Ca++-independent, shows maximal activity at pH 4, and represents a lysosomal-type PLA2. During the present period of grant support, we have shown that aiPLA2: 1) plays a major role in degradation of internalized alveolar DPPC; 2) provides substrate for the reacylation pathway of DPPC synthesis; 3) activity is regulated through protein-protein interactions with SP-A; 4) is localized in lung to lamellar bodies and lysosomes; 5) is activated by phosphorylation; and 6) its expression is developmental^ regulated and hormonally responsive. Specific Aim 1 will study models of under- and over-expression of aiPLA2 (knockout and transgenic mice) in order to evaluate the physiologic role of the enzyme in DPPC degradation and synthesis using isolated cells, the isolated perfused lung, and in vivo studies. Time-related (aging) changes in lung phospholipid content will be determined. Specific Aim 2 will evaluate the structure-function relationships for aiPLA2 activity utilizing