CDC14 PHOSPHORYLATION

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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 use our newly developed method for the isolation of GFP-tagged proteins to study phosphorylation of the phosphatase, Cdc14,required for cell cycle progression. GFP-tagged Cdc14, present at endogenous levels, was isolated from 16 L of yeast culture using magnetic beads coated with GFP Ab. Following digestion with trypsin (using various digestion conditions), several predicted sites of phosphorylation were analyzed by MS, MS/MS analyses and hypothesis driven mass spectrometry analysis.