Technologies Trageting Mitochondral Proteome
线粒体蛋白质组技术
基本信息
- 批准号:6992797
- 负责人:
- 金额:$ 9.99万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2005
- 资助国家:美国
- 起止时间:2005-09-01 至 2007-02-28
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
DESCRIPTION (provided by applicant): Mitochondria have a variety of essential responsibilities in cellular metabolism, e.g. the production of ATP through oxidative phosphorylation in aerobic cells as well as the initiation of the signal cascade leading to apoptosis. Alteration of the mitochondrial proteome and altered mitochondrial function have been implicated in a variety of degenerative diseases, heart disease, aging, and cancer. Because the mitochondria are a major source of endogenously generated reactive oxygen species (ROS), it is not surprising that they have been proposed to play a major role in aging. The free radical theory of aging predicts that oxidative damage to the mitochondria can lead to an amplifying effect whereby damaged mitochondria release more ROS, further increasing oxidative damage. Considering the progressive age-dependent functional changes of most cell types and tissues, an accurate profile of age-dependent mitochondria proteome is mandatory and will shed light on the molecular mechanisms and pathways of age-associated disease processes. To greatly increase the mitochondria proteome coverage, particularly toward the identification of post-translational modifications (PTMs), this project aims to develop and demonstrate a capillary gel electrophoresis (CGE)-based multidimensional protein separation platform, capable of providing significant analyte concentration and extremely high resolving power for handling complex protein mixtures prior to mass spectrometry detection. In comparison with conventional SDS-PAGE-based proteome approaches, the proposed CGE/nano-reversed-phase liquid chromatography proteome technology will be highly automated and offer robust and high throughput fractionation of whole proteins for top-down proteomics while avoiding analyte dilution and loss in an integrated platform. By coupling with a nanoscale trypsin membrane reactor, the ultrafast proteolytic digestion of proteins resolved and eluted from the CGE-based separations enables the combined top-down/bottom-up characterization of PTMs in mouse liver mitochondria as the model system in this study.
描述(由申请人提供):线粒体在细胞代谢中具有多种基本功能,例如,在需氧细胞中通过氧化磷酸化产生ATP以及启动导致细胞凋亡的信号级联。线粒体蛋白质组的改变和线粒体功能的改变与多种退行性疾病、心脏病、衰老和癌症有关。由于线粒体是内源性产生的活性氧(ROS)的主要来源,因此它们被认为在衰老中起主要作用也就不足为奇了。衰老的自由基理论预测,线粒体的氧化损伤可导致放大效应,由此受损的线粒体释放更多的ROS,进一步增加氧化损伤。 考虑到大多数细胞类型和组织的渐进性年龄依赖性功能变化,年龄依赖性线粒体蛋白质组的准确图谱是必需的,并将揭示年龄相关疾病过程的分子机制和途径。为了大大增加线粒体蛋白质组的覆盖率,特别是对翻译后修饰(PTM)的鉴定,本项目旨在开发和展示一种基于毛细管凝胶电泳(CGE)的多维蛋白质分离平台,能够提供显着的分析物浓度和极高的分辨率,用于在质谱检测之前处理复杂的蛋白质混合物。与传统的基于SDS-PAGE的蛋白质组方法相比,所提出的CGE/纳米反相液相色谱蛋白质组技术将高度自动化,并为自上而下的蛋白质组学提供完整蛋白质的稳健和高通量分级分离,同时避免分析物在集成平台中的稀释和损失。通过与纳米级胰蛋白酶膜反应器耦合,从基于CGE的分离中解析和洗脱的蛋白质的超快蛋白水解消化使得能够组合自上而下/自下而上表征作为本研究中的模型系统的小鼠肝线粒体中的PTM。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Brian M Balgley其他文献
Brian M Balgley的其他文献
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{{ truncateString('Brian M Balgley', 18)}}的其他基金
Development of a non-destructive molecular extraction platform for quantitative p
开发用于定量分析的无损分子提取平台
- 批准号:
8754932 - 财政年份:2014
- 资助金额:
$ 9.99万 - 项目类别:
Breast Cancer Proteomic via Laser-Free Microdissection and Gemini Technologies
通过无激光显微切割和 Gemini 技术进行乳腺癌蛋白质组学
- 批准号:
7290319 - 财政年份:2006
- 资助金额:
$ 9.99万 - 项目类别:
Identification of Differentially Expressed Proteomes in Human Astrocytomas
人星形细胞瘤中差异表达蛋白质组的鉴定
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7107560 - 财政年份:2006
- 资助金额:
$ 9.99万 - 项目类别:
Breast Cancer Proteomic via Laser-Free Microdissection and Gemini Technologies
通过无激光显微切割和 Gemini 技术进行乳腺癌蛋白质组学
- 批准号:
7137564 - 财政年份:2006
- 资助金额:
$ 9.99万 - 项目类别:
Validation and Quantification of FFPE Antigen Retrieval by Proteome Analysis
通过蛋白质组分析验证和定量 FFPE 抗原修复
- 批准号:
7137367 - 财政年份:2006
- 资助金额:
$ 9.99万 - 项目类别:
Integrated Top-Down/Bottom-Up Comprehensive Proteomics
集成的自上而下/自下而上的综合蛋白质组学
- 批准号:
7103624 - 财政年份:2004
- 资助金额:
$ 9.99万 - 项目类别:
Integrated Top-Down/Bottom-Up Comprehensive Proteomics
集成的自上而下/自下而上的综合蛋白质组学
- 批准号:
7017292 - 财政年份:2004
- 资助金额:
$ 9.99万 - 项目类别:
Integrated Top-Down/Bottom-Up Comprehensive Proteomics
集成的自上而下/自下而上的综合蛋白质组学
- 批准号:
6785714 - 财政年份:2004
- 资助金额:
$ 9.99万 - 项目类别:
ITP-Based Selective Enrichment of Low Abundance Proteins
基于 ITP 的低丰度蛋白质选择性富集
- 批准号:
6686685 - 财政年份:2003
- 资助金额:
$ 9.99万 - 项目类别:
ITP-Based Selective Enrichment of Low Abundance Proteins
基于 ITP 的低丰度蛋白质选择性富集
- 批准号:
6776989 - 财政年份:2003
- 资助金额:
$ 9.99万 - 项目类别:
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