CHARACTERIZATION OF NUCLEATION PROMOTING FACTOR PROTEIN COMPLEXES BY MASS SPEC

通过质谱对成核促进因子蛋白复合物进行表征

• 批准号: 8171327
• 负责人: Matthew D Welch
• 金额: $ 0.24万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171327
• 关键词: Actins; Cardiovascular Diseases; Cell division; Cell physiology; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytokinesis; Cytoskeletal Proteins; Cytoskeleton; Diagnosis; Disease; Embryonic Development; Endocytosis; Filament; Funding; Goals; Grant; Immune system; Infection; Inflammatory; Institution; Lead; Locomotion; Mammalian Cell; Maps; Mass Spectrum Analysis; Microfilaments; Neoplasm Metastasis; Phagocytosis; Phosphorylation; Phosphorylation Site; Play; Process; Proteins; Regulation; Research; Research Personnel; Resources; Role; Signal Transduction; Source; Time; Transmembrane Transport; United States National Institutes of Health; Virus Diseases; Work; Wound Healing; cell motility; microbial; novel strategies; polymerization; protein complex; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The actin cytoskeleton plays an essential role in basic cellular processes including locomotion, membrane transport, and cytokinesis. These processes are important for normal embryonic development, immune system function and tissue repair, and also contribute to illnesses such inflammatory disease, cardiovascular disease, cancer metastasis, and microbial infection. To enable actin function, cells precisely control the nucleation of actin polymerization and the organization of the resulting filaments. The goal of our work is to understand the function and regulation of one of the cell's key actin nucleating and organizing factors, the Arp2/3 complex. This protein complex polymerizes actin filaments into Y-branched networks that participate in lamellipodia protrusion, phagocytosis, endocytosis and membrane transport. Activation of the Arp2/3 complex requires proteins called nucleation promoting factors (NPFs), of which there are several in mammalian cells, each acting to direct the Arp2/3 complex to participate in a particular cellular process. NPFs are themselves regulated by other cytoskeletal proteins and signal transduction molecules that coordinate their activities in space and time. Despite the key importance of NPFs in regulating actin nucleation in cells, little is known about whether each exists in one or more protein complexes, what other proteins assemble into these complexes, and how the activity of these complexes is regulated by phosphorylation. The goal of this project is to use mass spectrometry to identify the protein components of NPF-containing protein complexes purified from mammalian cells, and to map potential regulatory phosphorylation sites in the components of these complexes. In the long term, these experiments will help elucidate how actin nucleation and organization is controlled in cells during processes such as cell locomotion, membrane transport, and cell division. Understanding these mechanisms may ultimately lead to new approaches to diagnose and treat inflammatory disease, cardiovascular disease, cancer metastasis, and bacterial and viral infection.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 肌动蛋白细胞骨架在运动、膜运输和胞质分裂等基本细胞过程中发挥着重要作用。 这些过程对于正常胚胎发育、免疫系统功能和组织修复非常重要，并且也会导致炎症性疾病、心血管疾病、癌症转移和微生物感染等疾病。为了实现肌动蛋白功能，细胞精确控制肌动蛋白聚合的成核和所得丝的组织。 我们工作的目标是了解细胞关键肌动蛋白成核和组织因子之一 Arp2/3 复合物的功能和调节。 这种蛋白质复合物将肌动蛋白丝聚合成 Y 分支网络，参与片状伪足突出、吞噬作用、内吞作用和膜运输。 Arp2/3 复合物的激活需要称为成核促进因子 (NPF) 的蛋白质，哺乳动物细胞中有多种此类蛋白质，每种蛋白质都可指导 Arp2/3 复合物参与特定的细胞过程。 NPF 本身受到其他细胞骨架蛋白和信号转导分子的调节，这些蛋白和信号转导分子在空间和时间上协调其活动。 尽管 NPF 在调节细胞中肌动蛋白成核方面至关重要，但人们对每种 NPF 是否存在于一个或多个蛋白质复合物中、哪些其他蛋白质组装到这些复合物中以及这些复合物的活性如何通过磷酸化进行调节仍知之甚少。该项目的目标是利用质谱法鉴定从哺乳动物细胞中纯化的含 NPF 的蛋白质复合物的蛋白质成分，并绘制这些复合物成分中潜在的调节磷酸化位点。 从长远来看，这些实验将有助于阐明细胞运动、膜运输和细胞分裂等过程中肌动蛋白成核和组织是如何控制的。 了解这些机制可能最终会带来诊断和治疗炎症性疾病、心血管疾病、癌症转移以及细菌和病毒感染的新方法。