ENZYMES MODULATING SECOND MESSENGERS IN NEUTROPHILS

描述(摘自申请者的摘要)：这是一份竞争性的 更新版，题为“调节中性粒细胞第二信使的酶”， 要求再寻找五年，10-14年。(改编自 申请人摘要)本提案的目的是定义 参与刺激的分子事件的确切序列 吞噬白细胞。从这些研究中获得的知识可能会 导致治疗传染病的新策略和 发炎。这个项目现在将重点放在两个截然不同但至关重要的 中性粒细胞刺激的反应：(1) Cofilin的去磷酸化/激活，以及(2)不同的作用 蛋白激酶C(PKC)同工酶在NADPH氧化酶活化中的作用 触发超氧化物生成的复合体。调查员和 合作者最近报告说，粘连蛋白经历了快速的 在刺激的中性粒细胞中去磷酸化/激活Cofilin是一种 重要的肌动蛋白解聚剂 调节肌动蛋白细胞骨架，特别是功能反应 中性粒细胞，如趋化、吞噬小体形成和脱颗粒。 此外，调查员已经提出证据表明Cofilin是 受一种新的磷酸化和去磷酸化循环调节 中性粒细胞。参与其中的激酶和磷酸酶的性质(S) 在这个循环中都是未知的。 具体地说，该项目重点关注四个未勘探地区(五个目标) 中性粒细胞的信号转导途径。这些是(1)识别 和鉴定蛋白激酶(AIM 1)和磷酸酶(AIMS 2) 催化档案的磷酸化和去磷酸化 中性粒细胞，(2)阐明触发 细胞刺激期间堆积的去磷酸化/激活(目标3)， (3)继续研究PKC在超氧化物生产中的作用 存在于中性粒细胞和中性粒细胞中的每一种纯化的PKC同工酶 它们的重组底物，即氧化酶亚基P47-Phox和P67- PHOX(目标4)，以及(4)监测超氧化物生成的组装 免疫荧光系统及其相互作用的表征 在这个复合体和细胞骨架之间(目标5)。共同本地化 特定同工酶的PKC与氧化物酶亚基将购买。这个 目标是在监管属性之间建立坚实的界限 分离的酶及其对刺激反应现象的控制 吞噬白细胞。

DESCRIPTION (Adapted from applicant's abstract): This is a competitive renewal entitled, "Enzymes modulating second messengers in neutrophils", requesting five more years of finding, for years 10-14. (adapted from the applicant's abstract) The objective of this proposal is to define the exact sequence of molecular events that are involved in the stimulation of phagocytic leukocytes. Knowledge gained from these studies will likely to lead to novel strategies for treating infectious diseases and inflammation. This project will now focus on two distinct but critical reactions in the stimulation of neutrophils: (1) the dephosphorylation/activation of cofilin, and (2) the role of different isozyme of protein kinase C (PKC) in the activation of the NADPH oxidase complex which triggers superoxide generation. The investigator and collaborators have recently reported that cofilin undergoes rapid dephosphorylation/activation in stimulated neutrophils Cofilin is an essential actin depolymerizing agent that is critically involved in regulating the actin cytoskeleton, particularly the functional responses of neutrophils, e.g., chemotaxis, phagosome formation and degranulation. Moreover, the investigator has presented evidence that cofilin is regulated by a novel cycle of phosphorylation and dephosphorylation in neutrophils. The nature of the kinases and phosphatase(s) that participate in this cycle are unknown. Specifically, this project focuses on four unexplored areas (five aims) in the signal transduction pathways of neutrophils. These are (1) identifying and characterizing the protein kinase (aim 1) and phosphatase (aims 2) that catalyze the phosphorylation and dephosphorylation of cofiling in neutrophils, (2) elucidating the regulatory mechanisms that trigger dephosphorylation/activation of cofiling during cell stimulation (aim 3), (3) continue investigating the role of PKC in superoxide production with each of the purified isozymes of PKC that are present in neutrophils and their recombinant substrates, i.e., the oxidase subunits p47-phox and p67- phox (aim 4), and (4) monitoring the assembly of the superoxide generating system by immunofluorescence techniques and characterizing interactions between this complex and the cytoskeleton (aim 5). Co-localization of specific isozymes of PKC with the oxidase subunits will be bought. The goal is to forge a solid line between the regulatory properties of the isolated enzymes and control of the stimulus-response phenomena in phagocytic leukocytes.