PHOSPHO-PROTEIN PHOSPHORYLATION SITE IDENTIFICATION METHODOLOGY

磷酸蛋白磷酸化位点识别方法

• 批准号: 8363747
• 负责人: ALMA L BURLINGAME
• 金额: $ 4.83万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363747
• 关键词: Affinity Chromatography; Chromatography; Complex Mixtures; Development; Fractionation; Funding; Grant; Ions; Mass Spectrum Analysis; Metals; Methodology; National Center for Research Resources; Peptides; Phase; Phosphorylation; Phosphorylation Site; Principal Investigator; Proteins; Reporting; Research; Research Infrastructure; Resources; Source; Techniques; Time; United States National Institutes of Health; base; cost; instrument; research and development; titanium dioxide

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Development of metal ion affinity chromatography (IMAC) and titanium dioxide (TiO2) based methodology for the isolation of phospho-peptides at the sub-picomole level. These enrichment techniques will be combined with off-line multidimensional chromatography fractionation prior to final analysis using reverse phase LC-MS/MS. Applications to determination of sites of phosphorylation on a variety of biologically important proteins, either in an isolated state or in the context of complex mixtures. (Additional effort and instrument time reported under Collaborative projects and other Technical Research and Development project.)

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 开发基于金属离子亲和色谱 (IMAC) 和二氧化钛 (TiO2) 的方法，用于在亚皮摩尔水平分离磷酸肽。 在使用反相 LC-MS/MS 进行最终分析之前，这些富集技术将与离线多维色谱分离相结合。适用于测定各种生物学上重要的蛋白质的磷酸化位点，无论是在分离状态还是在复杂混合物的情况下。 （在合作项目和其他技术研究与开发项目下报告了额外的工作量和仪器时间。）