title: Visualization and biochemical analyses of the emerging mammalian 14-3-3-phosphoproteome.
abstract: Hundreds of candidate 14-3-3-binding (phospho)proteins have been reported in publications that describe one interaction at a time, as well as high-throughput 14-3-3-affinity and mass spectrometry-based studies. Here, we transcribed these data into a common format, deposited the collated data from low-throughput studies in MINT (http://mint.bio.uniroma2.it/mint), and compared the low- and high-throughput data in VisANT graphs that are easy to analyze and extend. Exploring the graphs prompted questions about technical and biological specificity, which were addressed experimentally, resulting in identification of phosphorylated 14-3-3-binding sites in the mitochondrial import sequence of the iron-sulfur cluster assembly enzyme (ISCU), cytoplasmic domains of the mitochondrial fission factor (MFF), and endoplasmic reticulum-tethered receptor expression-enhancing protein 4 (REEP4), RNA regulator SMAUG2, and cytoskeletal regulatory proteins, namely debrin-like protein (DBNL) and kinesin light chain (KLC) isoforms. Therefore, 14-3-3s undergo physiological interactions with proteins that are destined for diverse subcellular locations. Graphing and validating interactions underpins efforts to use 14-3-3-phosphoproteomics to identify mechanisms and biomarkers for signaling pathways in health and disease.
authors: MRC Protein Phosphorylation Unit, College of Life Sciences, University of Dundee, James Black Centre, Dow Street, Dundee DD1 5EH, Scotland, UK.
authors: Catherine
authors: C
authors: Johnson
authors: Michele
authors: M
authors: Tinti
authors: Nicola T
authors: NT
authors: Wood
authors: David G
authors: DG
authors: Campbell
authors: Rachel
authors: R
authors: Toth
authors: Fanny
authors: F
authors: Dubois
authors: Kathryn M
authors: KM
authors: Geraghty
authors: Barry H C
authors: BH
authors: Wong
authors: Laura J
authors: LJ
authors: Brown
authors: Jennifer
authors: J
authors: Tyler
authors: Aurélie
authors: A
authors: Gernez
authors: Shuai
authors: S
authors: Chen
authors: Silvia
authors: S
authors: Synowsky
authors: Carol
authors: C
authors: MacKintosh
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D015415:Biomarkers / Q000378:metabolism; D030562:Databases, Protein; D057809:HEK293 Cells; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D048948:14-3-3 Proteins; D015415:Biomarkers; D010750:Phosphoproteins; D011994:Recombinant Proteins