Abstract
14-3-3 proteins regulate many intracellular processes and their ability to bind in subtly different fashions to their numerous partner proteins provides attractive drug-targeting points for a range of diseases. Schnurri-3 is a suppressor of mouse bone formation and a candidate target for novel osteoporosis therapeutics, and thus it is of interest to determine whether it interacts with 14-3-3. In this work, potential 14-3-3 interaction sites on mammalian Schnurri-3 were identified by anin silico analysis of its protein sequence. Using fluorescence polarization, isothermal titration calorimetry and X-ray crystallography, it is shown that synthetic peptides containing either phosphorylated Thr869 or Ser542 can indeed interact with 14-3-3, with the latter capable of forming an interprotein disulfide bond with 14-3-3σ: a hitherto unreported phenomenon.
Highlights
14-3-3 proteins are a family of regulatory adaptor proteins with an interactome currently estimated to include almost 500 binding partners (Aitken, 2006)
Tens of crystal structures of 14-3-3 proteins bound to these motifs in the form of synthetic phosphorylated peptides have been deposited in the PDB (Sluchanko, 2018), and together they provide an important resource that can be exploited for the understanding and modulation of 14-3-3 protein–protein interactions (PPIs)
We performed an in silico analysis and found 18 potential 14-3-3 interaction sites in human SHN3, only two of which were conserved in the mouse sequence
Summary
14-3-3 proteins are a family of regulatory adaptor proteins with an interactome currently estimated to include almost 500 binding partners (Aitken, 2006). They facilitate cellular processes such as signal transduction, cellular trafficking, apoptosis and cell-cycle regulation (Hermeking & Benzinger, 2006; Mackintosh, 2004; Johnson et al, 2011). The binding of 14-3-3 proteins to their protein partners occurs via the recognition of specific consensus motifs that includes a phosphorylated serine or threonine. The interaction of SHN3 with kinases opens up the possibility that its function is regulated by phosphorylation and binding to 14-3-3 proteins. We observed that the motif built around Ser542 in human SHN3 can become covalently associated with 14-3-3 via an interprotein disulfide bond
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 callMore From: Acta crystallographica. Section F, Structural biology communications
Disclaimer: 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.
