FINDING NEK2 KINASE AUTO AND SUBSTRATE PHOSPHORYLATION SITES IN HUMAN CELLS

寻找人类细胞中的 NEK2 激酶自动和底物磷酸化位点

• 批准号: 8169805
• 负责人: Jack Taunton
• 金额: $ 0.53万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-12 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169805
• 关键词: Cells; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Epitopes; Event; Funding; Grant; Human; Institution; Link; Mediating; Metabolic Pathway; Methods; Phosphorylation; Phosphorylation Site; Phosphotransferases; Protein Kinase; Proteins; Research; Research Personnel; Resistance; Resources; Sampling; Signal Transduction; Source; United States National Institutes of Health; autophosphorylation-dependent multifunctional protein kinase; cell behavior; inhibitor/antagonist; interest; mutant; small molecule

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. Protein kinases are a diverse set of regulatory enzymes responsible for signaling in a majority of metabolic pathways and cell behaviors. Kinases control signaling events within the cell through phosphorylation of substrate proteins. Identification of a given kinases' substrate proteins and their specific phosphorylation sites is essential to understanding kinases mediated signaling events. However, there is no general or trivial method for such identifications. We are investigating a method of elucidating kinase phosphorylation sites using small molecule inhibitors of a specific kinase, Nek2, as well as an inhibitor resistant mutant of Nek2 kinase. The method involves over-expressing the kinase of interest (Nek2) in human cells with an epitope tag that can be used to purify Nek2 and associated proteins from these cells. Mass Spec analysis can then be used to identify phosphorylation sites on these purified proteins. By making a quantitative comparison between samples treated with out Nek2 inhibitor and samples expressing the inhibitor resistant Nek2 mutant we can determine phosphorylation sites specifically linked to Nek2 kinase activity.

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