Affinity methods for phosphorylation-dependent interactions.

Greg Moorhead; Carol MacKintosh

doi:10.1385/1-59259-762-9:469

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Affinity methods for phosphorylation-dependent interactions.

Greg Moorhead·Carol MacKintosh

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Abstract

14-3-3s are a highly conserved protein family that exert many regulatory roles in eukaryotic cells by binding to phosphopeptide motifs in diverse target proteins. Here, we describe 14-3-3 affinity binding procedures that can be used to purify and identify 14-3-3-binding phosphoproteins; monitor how their phosphorylation and 14-3-3 binding is regulated by extracellular stimuli; define the functional effects of 14-3-3s on individual targets; and identify relevant protein phosphatases and kinases. In principle, these methods could be adapted to characterize other types of protein-protein interaction that depend on covalent modification of target sites.

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14-3-3 Proteins Amino Acid Sequence Animals Chromatography, Affinity Molecular Sequence Data Phosphopeptides Phosphorylation Protein Binding Tyrosine 3-Monooxygenase Journal Article Review

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Greg Moorhead, Carol MacKintosh. 2004. Affinity methods for phosphorylation-dependent interactions.. https://doi.org/10.1385/1-59259-762-9%3A469

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