Intramolecular domain dynamics regulate synaptic MAGUK protein interactions
- Charité-Universitätsmedizin Berlin, Germany
- Freie Universität Berlin, Germany
- Helmholtz-Zentrum Berlin für Materialien und Energie, Germany
Figures
Figure 1 with 2 supplements
PDZ3 ligand-induced dynamics in the PDZ3-SH3-GK module facilitate oligomerisation.
(A) Schematic representation of the PSD-95 domain organisation. PSD-95 contains three PDZ domains followed by a SH3-GK domain tandem. The PSG module (PDZ3-SH3-GK) is common to the MAGUK protein …
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Figure 1—source data 1
Source data for Figure 1C,D.
- https://doi.org/10.7554/eLife.41299.003
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Figure 1—source data 2
Source data for Figure 1E.
- https://doi.org/10.7554/eLife.41299.004
Figure 1—figure supplement 1
FACS plots for Figure 1C,D.
(A, B) Gating strategy and representative dot plots of the PSD-95 oligomerisation assay as shown in Figure 1C,D. Untransfected cells or cells transfected with the indicated constructs were harvested …
Figure 1—figure supplement 2
Supplement for Figure 1D.
(A) PSD-95 constructs consisting of the PDZ3-SH3 domains (PS) were coexpressed together with either an SH3-GK domain construct, or a GK domain construct. As a comparison PDZ3-SH3 L460P was …
Figure 2
Identification of interactors that differentially bind to PSD-95 C-terminal domains.
(A) Schematic representation of the quantitative mass spectrometry experiment to identify PSD-95 GK domain interactors from crude rat synaptosomes by GST pull-down of bacterially expressed GST-GK or …
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Figure 2—source data 1
Source data for Figure 2B.
- https://doi.org/10.7554/eLife.41299.008
Figure 3
The heterotrimeric G protein subunit Gnb5 is a novel PSD-95 interactor.
(A) GST pull-down from crude synaptosomal proteins (comparable amounts of GST tagged proteins observable by Coomassie, upper panel) enabled comparison of Gnb5 binding to the GK domain alone versus …
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Figure 3—source data 1
Source data for Figure 3A,B.
- https://doi.org/10.7554/eLife.41299.010
Figure 4
Gnb5 - PSD-95 complex formation is regulated by CRIPT-derived PDZ3 ligand binding.
For this figure, western blots shown on the left side are representative examples of three independent experiments; the corresponding quantification of coIP band intensities from these three …
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Figure 4—source data 1
Source data for Figure 4.
- https://doi.org/10.7554/eLife.41299.012
Figure 5
GK domain interactions are differentially regulated.
For this figure, western blots shown on the left side are representative examples of three independent experiments; the corresponding quantification of coIP band intensities from these three …
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Figure 5—source data 1
Source data for Figure 5.
- https://doi.org/10.7554/eLife.41299.014
Figure 6
Graphical Summary.
(A) PSD-95 C-terminal domains (PSG module) functionally cooperate and regulate homotypic and heterotypic complex formation. We propose that CRIPT-derived PDZ3 ligand binding to the PDZ3 domain …
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.41299.016
