Visualization and Biochemical Analyses of the Emerging Mammalian 14-3-3-Phosphoproteome

• DOI: 10.1074/mcp.m110.005751
• Publication Date: 2011-07-04T02:43:06Z
• Authors: Catherine Johnson; Michele Tinti; Nicola T. Wood; David G. Campbell; Rachel Toth; Fanny Dubois; Kathryn M. Geraghty; Barry H.C. Wong; Laura J. Brown; Jennifer Tyler; Aurélie Gernez; Shuai Chen; Silvia Synowsky; Carol MacKintosh
• Keywords: 0301 basic medicine; 570; Binding Sites; Research; Phosphoproteins; Mass Spectrometry; Recombinant Proteins; Mitochondria; Protein Structure, Tertiary; Mice; 03 medical and health sciences; HEK293 Cells; 14-3-3 Proteins; Animals; Humans; Databases, Protein; Biomarkers; Protein Binding; Signal Transduction

Hundreds of candidate 14-3-3-binding (phospho)proteins have been reported in publications that describe one interaction at a time, as well high-throughput 14-3-3-affinity and mass spectrometry-based studies. Here, we transcribed these data into common format, deposited the collated from low-throughput studies MINT (http://mint.bio.uniroma2.it/mint), compared low- VisANT graphs are easy to analyze extend. Exploring prompted questions about technical biological specificity, which were addressed experimentally, resulting identification phosphorylated sites mitochondrial import sequence iron-sulfur cluster assembly enzyme (ISCU), cytoplasmic domains fission factor (MFF), endoplasmic reticulum-tethered receptor expression-enhancing protein 4 (REEP4), RNA regulator SMAUG2, cytoskeletal regulatory proteins, namely debrin-like (DBNL) kinesin light chain (KLC) isoforms. Therefore, 14-3-3s undergo physiological interactions with proteins destined for diverse subcellular locations. Graphing validating underpins efforts use 14-3-3-phosphoproteomics identify mechanisms biomarkers signaling pathways health disease.