PHOSPHOPROTEOMICS OF EPISTATICALLY INTERACTING KINASES AND PHOSPHATASES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们之前完成了对酵母中物理蛋白质相互作用的大规模分析，结果鉴定了500多个蛋白质复合体(自然440,637)。最近，我们与Kevin Shokat教授合作完成了上位性相互作用基因的筛选，并发现一对相互作用的基因与物理相互作用密切相关，特别是在一对激酶/磷酸酶及其底物蛋白之间。在这个项目中，亲和标记的潜在底物蛋白将从酵母中纯化，消化成多肽，并使用Beltrao等人的方法富集磷酸肽(PLoS Biol.7(6)：e1000134)。浓缩组分将用LC MS进行分析。

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. We previously completed a large-scale analysis of physical protein interactions in yeast resulting in the identification of over 500 protein complexes (Nature 440, 637). More recently, we completed a screen for epistatically interacting genes in collaboration with Prof. Kevan Shokat, and have found that pairs of genes interacting positively correlate strongly with physical interactions, in particular between pairs of kinases/phosphatases and their substrate proteins. In this project, affinity tagged potential substrate proteins will be purified from yeast, digested into peptides, and enriched for phosphopeptides using the method of Beltrao et al., (PLoS Biol. 7(6):e1000134). Enriched fractions will be analyzed by LCMS.