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.
Highlights
14-3-3s interact with hundreds of phosphoproteins inside all eukaryotic cells, including mammalian proteins that are deregulated in diabetes, cancer, platelet disorders, viral infections, and neurological disorders [1]
The collective data highlighted that 14-3-3 dimers frequently engage with two phosphorylated motifs on their targets, and phosphorylated 14-3-3-binding sites fall into subtypes that overlap with the specificities of different basophilic protein kinases such as PKA, Akt/PKB, p90RSK, PKCs, and AMPK
As well as the low-throughput studies, high-throughput proteomics experiments have identified large pools of proteins that display affinity for 14-3-3s in extracts of human cells, rodent cells and tissues, bovine sperm, hydra, Saccharomyces cerevisiae, and plants [2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26]. These experiments used a variety of 14-3-3 capture and release strategies, and were performed over a 6-year period with changing state-ofthe-art in mass spectrometry
Summary
14-3-3s interact with hundreds of phosphoproteins inside all eukaryotic cells, including mammalian proteins that are deregulated in diabetes, cancer, platelet disorders, viral infections, and neurological disorders [1]. The collective data highlighted that 14-3-3 dimers frequently engage with two phosphorylated motifs on their targets, and phosphorylated 14-3-3-binding sites fall into subtypes that overlap with the specificities of different basophilic protein kinases such as PKA, Akt/PKB, p90RSK, PKCs, and AMPK. These specificities for 14-3-3s are consistent with the emerging roles for 14-3-3s in integrating cellular responses to insulin, growth factors, and nutrients [2,3,4]. This is a pity because these data could be a valuable reference for ongoing projects to map dynamic changes in the 14-3-3-phosphoproteome in response to stimuli, drugs and disease
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
