Fuzzy supertertiary interactions within PSD-95 enable ligand binding
- Clemson University, United States
- Stony Brook University, United States
- Heinrich-Heine-University Düsseldorf, Germany
- Heinrich Heine University Düsseldorf, Germany
Abstract
The scaffold protein PSD-95 links postsynaptic receptors to sites of presynaptic neurotransmitter release. Flexible linkers between folded domains in PSD-95 enable a dynamic supertertiary structure. Interdomain interactions within the PSG supramodule, formed by PDZ3, SH3 and Guanylate Kinase domains, regulate PSD-95 activity. Here we combined Discrete Molecular Dynamics and single molecule FRET to characterize the PSG supramodule, with time resolution spanning picoseconds to seconds. We used a FRET network to measure distances in full-length PSD-95 and model the conformational ensemble. We found that PDZ3 samples two conformational basins, which we confirmed with disulfide mapping. To understand effects on activity, we measured binding of the synaptic adhesion protein neuroligin. We found that PSD-95 bound neuroligin well at physiological pH while truncated PDZ3 bound poorly. Our hybrid structural models reveal how the supertertiary context of PDZ3 enables recognition of this critical synaptic ligand.
Data availability
Datasets of FRET values from smTIRF have been uploaded as Figure 4-Source Data 1. The MATLAB scripts used to analyze smTIRF data have been uploaded as Source Code 1. Datasets from Confocal Microscopy with Multiparameter Fluorescence Detection (Raw, MFD Bursts, TCSPC, PDA, and FCS), along with structures used in generating simulated distances and DMD screening, are available at Zenodo (DOI: 10.5281/zenodo.6983428). Datasets from Discrete Molecular Dynamics are available at https://dlab.clemson.edu/research/PSD95-PSG/.
Article and author information
Author details
Funding
National Institute of Mental Health (MH081923)
- Mark E Bowen
National Institute of General Medical Sciences (GM130451)
- Hugo Sanabria
National Institute of General Medical Sciences (GM119691)
- Feng Ding
National Science Foundation (MCB1749778)
- Hugo Sanabria
National Science Foundation (CBET1553945)
- Feng Ding
European Research Council (671208)
- George L Hamilton
- Claus AM Seidel
- Hugo Sanabria
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Hamilton et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Fuzzy supertertiary interactions within PSD-95 enable ligand binding
eLife 11:e77242.
https://doi.org/10.7554/eLife.77242
Further reading
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By influencing calcium homeostasis, local protein synthesis and the endoplasmic reticulum, a small protein called Rab10 emerges as a crucial cytoplasmic regulator of neuropeptide secretion.
