Molecular Structure of the Phagocyte NADPH Oxidase

吞噬细胞 NADPH 氧化酶的分子结构

基本信息

批准号：
7216290
负责人：
MARK T QUINN
金额：
$ 29.49万
依托单位：
MONTANA STATE UNIVERSITY - BOZEMAN
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-09-30 至 2009-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The generation of 02- by phagocytic cells requires the assembly of a membrane-bound complex known as the NADPH oxidase, which is composed of an integral plasma membrane flavocytochrome b and four cytosolic proteins (p40 phox, p47 phox, p67 phox, and Rac2). Although progress has been made, there is limited information about how each of these proteins regulate formation of the active NADPH oxidase complex and, similarly, what turns off the system. In the previous grant period, we identified a number of protein-protein interactions among the NADPH oxidase protein components and investigated how these interactions related to assembly of the active 02- generating system. In this renewal application, we propose studies to investigate how NADPH oxidase assembly is regulated. Based on studies performed under the previous grant period, studies by others on the phagocyte NADPH oxidase, and preliminary work described here, we hypothesize that neutrophil capacity for oxidase assembly and oxidase assembly per se are regulated at several different levels, including transcription and biosynthesis of the phox proteins, molecular assembly of the active system, and lateral organization in plasma membrane domains. Consequently, appropriate NADPH oxidase activity results from integration of these multiple levels of regulation. Because of the complex nature of the oxidase, we propose to address this hypothesis by focusing this project primarily on the role of the cytosolic phox factors in oxidase regulation. Specifically, this proposal describes strategies for: 1) characterizing the promoter for p67 phox and determining what transcription factors regulate p67 phox transcription; 2) characterizing cytosolic phox protein gene expression; 3) analyzing biosynthesis of the cytosolic phox proteins to determine relative stability of these proteins and kinetics of protein turnover; 4) identifying and characterizing flavocytochrome b domains that bind p67 phox and Rac; and 5) develop cell lines expressing GFP-phox fusion proteins for use analysis of the intermolecular interactions between cytosolic oxidase proteins and the oxidase complex in vivo. The results of these studies will further our understanding of how distinct levels of regulatory input are integrated to facilitate functional assembly and activation of the phagocyte NADPH oxidase.

描述（由申请人提供）：由吞噬细胞生成02-需要组装一个称为NADPH氧化酶的膜结合的络合物，该复合酶由整合性质膜B和四个细胞质蛋白组成（p40 Phox，P47 Phox，P47 Phox，P67 Phox和Racox，p47 Phox，p67 Phox和Rac2）。尽管取得了进展，但有关这些蛋白质中的每一种如何调节活性NADPH氧化酶复合物的形成的信息有限，并且类似地关闭了系统。在上一个赠款期间，我们确定了NADPH氧化酶蛋白成分之间的许多蛋白质 - 蛋白质相互作用，并研究了这些相互作用如何与活性02-生成系统组装相关。在此续签应用中，我们建议研究如何调节NADPH氧化酶的组装。 Based on studies performed under the previous grant period, studies by others on the phagocyte NADPH oxidase, and preliminary work described here, we hypothesize that neutrophil capacity for oxidase assembly and oxidase assembly per se are regulated at several different levels, including transcription and biosynthesis of the phox proteins, molecular assembly of the active system, and lateral organization in plasma membrane domains.因此，适当的NADPH氧化酶活性是由这些多个调节水平的整合而产生的。由于氧化酶的复杂性质，我们建议通过将该项目重点集中在胞质Phox因子在氧化酶调节中的作用上来解决这一假设。具体而言，该建议描述了以下方面的策略：1）表征p67 phox的启动子并确定哪些转录因子调节p67 phox转录； 2）表征胞质Phox蛋白基因表达； 3）分析胞质Phox蛋白的生物合成，以确定这些蛋白质的相对稳定性和蛋白质转换的动力学； 4）识别和表征结合p67 phox和RAC的黄素色素B域； 5）开发表达GFP-Phox融合蛋白的细胞系，用于使用体内胞质氧化酶蛋白和氧化酶复合物之间分子间相互作用的分析。这些研究的结果将进一步了解不同水平的调节输入水平以促进功能组装和吞噬细胞NADPH氧化酶的激活。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Superoxide production in the vasculature of lipopolysaccharide-treated rats and pigs.

脂多糖处理的大鼠和猪的脉管系统中超氧化物的产生。

DOI：
10.1097/01.shk.0000054374.88889.37
发表时间：
2003
期刊：
Shock (Augusta, Ga.)
影响因子：
0
作者：
Javesghani,Danesh;Hussain,SabahNA;Scheidel,Jonathan;Quinn,MarkT;Magder,SheldonA
通讯作者：
Magder,SheldonA

Inhibition of peroxynitrite-mediated tyrosine nitration by a novel pyrrolopyrimidine antioxidant.

新型吡咯并嘧啶抗氧化剂抑制过氧亚硝酸盐介导的酪氨酸硝化。