ENZYMES MODULATING SECOND MESSENGERS IN NEUTROPHILS

描述（改编自申请人的摘要）：这是一个竞争性的 标题为“中性粒细胞中调节第二信使的酶”的更新， 要求再找五年，10-14年。（改编自The 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 吞噬性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， 炎症这个项目现在将集中在两个不同的，但关键的 刺激中性粒细胞的反应：（1） cofilin的去磷酸化/活化，和（2）不同的 蛋白激酶C（PKC）同工酶在NADPH氧化酶活化中的作用 引发超氧化物生成的复合物。研究者和 合作者最近报道，cofilin经历快速 Cofilin在刺激的嗜中性粒细胞中的去磷酸化/活化是一种 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，例如，趋化性、吞噬体形成和脱粒。 此外，研究人员提出证据表明，cofilin是 由磷酸化和去磷酸化的新循环调节， 中性粒细胞激酶和磷酸酶的性质， 在这个循环中是未知的。 具体而言，该项目侧重于四个未探索的领域（五个目标）， 中性粒细胞的信号转导途径。（1）识别 并表征蛋白激酶（AIM 1）和磷酸酶（AIM 2） 催化共折叠的磷酸化和去磷酸化， 中性粒细胞，（2）阐明触发的调节机制 细胞刺激期间共填充的去磷酸化/活化（AIM 3）， (3)继续研究PKC在超氧化物产生中的作用， 存在于中性粒细胞中的每种纯化的PKC同工酶， 它们的重组底物，即，氧化酶亚基p47-phox和p67- phox（aim 4），和（4）监测超氧化物生成的组装 通过免疫荧光技术和表征相互作用 该复合物和细胞骨架之间的关系（目的5）。的共定位 将购买具有氧化酶亚基的PKC的特异性同工酶。的 目标是在监管属性之间建立一条坚实的界限， 分离的酶和控制的刺激反应现象， 吞噬性白细胞

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.