M-Track: detecting short-lived protein-protein interactions in vivo.
We developed a protein-proximity assay in yeast based on fusing a histone lysine methyltransferase onto a bait and its substrate onto a prey. Upon binding, the prey is stably methylated and detected by methylation-specific antibodies. We applied this approach to detect varying interaction affinities among proteins in a mitogen-activated protein ... kinase pathway and to detect short-lived interactions between protein phosphatase 2A and its substrates that have so far escaped direct detection.
Mesh Terms:
Cell Cycle Proteins, Histone-Lysine N-Methyltransferase, Humans, Methylation, Mitogen-Activated Protein Kinases, Protein Interaction Mapping, Protein Phosphatase 2, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Substrate Specificity, TOR Serine-Threonine Kinases, Tacrolimus Binding Proteins
Cell Cycle Proteins, Histone-Lysine N-Methyltransferase, Humans, Methylation, Mitogen-Activated Protein Kinases, Protein Interaction Mapping, Protein Phosphatase 2, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Substrate Specificity, TOR Serine-Threonine Kinases, Tacrolimus Binding Proteins
Nat. Methods
Date: Jun. 01, 2012
PubMed ID: 22581371
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