Bidirectional protein–protein interactions control liquid–liquid phase separation of PSD-95 and its interaction partners
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The organization of the postsynaptic density (PSD), a protein-dense semi-membraneless organelle, is mediated by numerous specific protein–protein interactions (PPIs) which constitute a functional postsynapse. The PSD protein 95 (PSD-95) interacts with a manifold of proteins, including the C-terminal of transmembrane AMPA receptor (AMPAR) regulatory proteins (TARPs). Here, we uncover the minimal essential peptide responsible for the Stargazin (TARP-γ2)-mediated liquid–liquid phase separation (LLPS) formation of PSD-95 and other key protein constituents of the PSD. Furthermore, we find that pharmacological inhibitors of PSD-95 can facilitate the formation of LLPS. We found that in some cases LLPS formation is dependent on multivalent interactions, while in other cases short, highly charged peptides are sufficient to promote LLPS in complex systems. This study offers a new perspective on PSD-95 interactions and their role in LLPS formation, while also considering the role of affinity over multivalency in LLPS systems.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 103808 |
| Tidsskrift | iScience |
| Vol/bind | 25 |
| Udgave nummer | 2 |
| Antal sider | 26 |
| ISSN | 2589-0042 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
We would like to thank Prof. Mingjie Zhang and Prof. Daniel Choquet for providing plasmids. We would also like to thank Simon Erlendsson for fruitful discussions and Nabeela Khadim for technical assistance. We also acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen, and the cOpenNMR facility (Novo Nordisk Foundation, NNF18OC0032996) at the Department of Biology, University of Copenhagen. We also kindly acknowledge the team at FIDABio (Copenhagen, Denmark) for their great help and assistance. D.E.O. thanks the Carlsberg Foundation (grant nr. CF14-0284 ) and the Novo Nordisk Foundation (grant nr. 12953 ) for support.
Funding Information:
We would like to thank Prof. Mingjie Zhang and Prof. Daniel Choquet for providing plasmids. We would also like to thank Simon Erlendsson for fruitful discussions and Nabeela Khadim for technical assistance. We also acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen, and the cOpenNMR facility (Novo Nordisk Foundation, NNF18OC0032996) at the Department of Biology, University of Copenhagen. We also kindly acknowledge the team at FIDABio (Copenhagen, Denmark) for their great help and assistance. D.E.O. thanks the Carlsberg Foundation (grant nr. CF14-0284) and the Novo Nordisk Foundation (grant nr. 12953) for support. N.R.C, K.S, and K.L.M conceived the project. N.R.C performed protein expression and purification, conducted, and analyzed data from circular dichroism, size exclusion chromatography, fluorescence polarization, flow-induced dispersion analysis, SDS-PAGE sedimentation assays, fluorescence confocal microscopy, and celluSPOT experiments. C.P.P and K.T.E performed and analyzed the NMR experiments. N.R.C. J.N.P, and D.E.O. performed and analyzed the size exclusion chromatography multi-angle light scattering experiments. A.T.S and N.R.C designed multimeric peptides. V.S designed, synthesized, and printed the celluSPOT arrays. M.V.M did LC-MS and UPLC. N.R.C, K.L.M, and K.S wrote the article with contributions from all authors. A patent application has been filed with regards to multimeric PSD-95 inhibitors, see WO2021/176,082.
Publisher Copyright:
© 2022 The Authors
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