IDENTIFICATION OF NEW EFFECTORS FOR RAS FAMILY GTPASES AND ELUCIDATION OF THEIR

RAS 家族 GTP 酶新效应子的鉴定及其阐明

• 批准号: 7957384
• 负责人: FRANK PATRICK MCCORMICK
• 金额: $ 0.92万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957384
• 关键词: Affinity Chromatography; Cells; Colon Carcinoma; Computer Retrieval of Information on Scientific Projects Database; Coomassie blue; Family; Funding; GTP Binding; Grant; Guanosine Triphosphate Phosphohydrolases; Institution; Mass Spectrum Analysis; Methodology; Play; Proteins; Ras Signaling Pathway; Research; Research Personnel; Resources; Role; Sequence Homology; Source; Staining method; Stains; United States National Institutes of Health; base; cancer cell; cell growth; gel electrophoresis; insight; member; protein complex; tumorigenesis

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 Ras signaling pathway plays a crucial role in regulating cell growth and tumorigenesis. Ras is a member of a sub-family of related GTPases that functions by regulating a variety of downstream targets of effectors. Based on sequence homologies in the Ras Association domain, we have identified serveral new Ras family GTPase effectors and confirmed that these new proteins can interact with Ras when in its active, GTP-bound state. Very little, if anything, is known about these new Ras effectors. As a way of getting some insight into their function and mechanism of action, we are trying to identify the proteins they interact with within the cell. We have used the Tandem Affinity Purification (TAP) methodology to express and purify from colon cancer cells TAP-tagged versions of these new Ras effectors. After separation of the protein complexes by gel electrophoresis and coomassie blue staining, we can visualize several co-purifying protein bands that we would now like to identify by mass spectrometry.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 RAS信号通路在调节细胞生长和肿瘤发生中起着至关重要的作用。 RAS是相关GTPase的子家庭的成员，该子族通过调节各种下游效应子的靶标而起作用。 基于RAS关联域中的序列同源性，我们已经确定了Serveral New RAS家族GTPase效应子，并确认这些新蛋白在其活跃的GTP结合状态时可以与RAS相互作用。 对于这些新的RAS效应子，几乎没有什么了解（如果有的话）。 为了了解其功能和作用机理的某种见解，我们正在尝试识别它们在细胞中与之相互作用的蛋白质。 我们已经使用了串联亲和力纯化（TAP）方法来表达和纯化这些新RAS效应子的结肠癌细胞TAP标签版本。 通过凝胶电泳和coomassie蓝色染色将蛋白质复合物分离后，我们可以可视化几个共纯化的蛋白质带，我们现在想通过质谱法鉴定。