title: M-Track: detecting short-lived protein-protein interactions in vivo.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Cell Biology, Max F. Perutz Laboratories, University of Vienna, Austria.
authors.forename: Aurora
authors.initials: A
authors.lastname: Zuzuarregui
authors.forename: Thomas
authors.initials: T
authors.lastname: Kupka
authors.forename: Bhumika
authors.initials: B
authors.lastname: Bhatt
authors.forename: Ilse
authors.initials: I
authors.lastname: Dohnal
authors.forename: Ingrid
authors.initials: I
authors.lastname: Mudrak
authors.forename: Christina
authors.initials: C
authors.lastname: Friedmann
authors.forename: Stefan
authors.initials: S
authors.lastname: Schüchner
authors.forename: Ingrid E
authors.initials: IE
authors.lastname: Frohner
authors.forename: Gustav
authors.initials: G
authors.lastname: Ammerer
authors.forename: Egon
authors.initials: E
authors.lastname: Ogris
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D054648:Protein Phosphatase 2 / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013379:Substrate Specificity; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D022021:Tacrolimus Binding Proteins / Q000378:metabolism
chemical_list: C071246:CDC55 protein, S cerevisiae; D018797:Cell Cycle Proteins; D029701:Saccharomyces cerevisiae Proteins; D011495:Histone-Lysine N-Methyltransferase; D058570:TOR Serine-Threonine Kinases; D020928:Mitogen-Activated Protein Kinases; D054648:Protein Phosphatase 2; D022021:Tacrolimus Binding Proteins