Synaptic Targeting and Function of SAPAPs Mediated by Phosphorylation-Dependent Binding to PSD-95 MAGUKs

Zhu, Jinwei; Zhou, Qingqing; Shang, Yuan; Li, Hao; Peng, Mengjuan; Ke, Xiao; Weng, Zhuangfeng; Zhang, Rongguang; Huang, Xuhui; Li, Shawn S.‐C.; Feng, Guoping; Lu, Youming; Zhang, Mingjie

doi:10.1016/j.celrep.2017.11.107

Cited by 52 publications

(61 citation statements)

References 67 publications

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“…To obtain soluble and well behaving GKAP and Shank3, we removed part of the sequences of the two proteins (see Figure 1A and STAR Methods for details). The binding affinities between PSD-95 and GKAP, between GKAP and Shank3, and between Shank3 and Homer3 were essentially the same as the bindings investigated using the isolated domains as reported previously (Zeng et al, 2016b;Zhu et al, 2017) . We also confirmed that Homer3 is a homo-tetramer (Hayashi et al, 2006(Hayashi et al, , 2009.…”

Section: Psd Condensates Formation By Mixing Major Psd Scaffold Proteinssupporting

confidence: 80%

“…To obtain soluble and well behaving GKAP (aka SAPAP1 or DLGAP1) and Shank3, we removed part of the sequences of the two proteins. Specifically, we removed part of the sequences on GKAP connecting three PSD-95 GK-binding repeats (GBRs) (Kim et al, 1997;Zhu et al, 2017) and a C-terminal extended PDZ-binding motif (PBM) required for stable Shank binding (Zeng et al, 2016b). Shank3 is a very large scaffold protein that is difficult to obtain a highly purified, well behaving full-length recombinant protein.…”

Section: Methods Details Protein Expression and Purificationmentioning

confidence: 99%

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Reconstituted Postsynaptic Density as a Molecular Platform for Understanding Synapse Formation and Plasticity

Zeng

Chen

Guan

et al. 2018

Cell

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354

407

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Synapses are semi-membraneless, protein-dense, sub-micron chemical reaction compartments responsible for signal processing in each and every neuron. Proper formation and dynamic responses to stimulations of synapses, both during development and in adult, are fundamental to functions of mammalian brains, although the molecular basis governing formation and modulation of compartmentalized synaptic assemblies is unclear. Here, we used a biochemical reconstitution approach to show that, both in solution and on supported membrane bilayers, multivalent interaction networks formed by major excitatory postsynaptic density (PSD) scaffold proteins led to formation of PSD-like assemblies via phase separation. The reconstituted PSD-like assemblies can cluster receptors, selectively concentrate enzymes, promote actin bundle formation, and expel inhibitory postsynaptic proteins. Additionally, the condensed phase PSD assemblies have features that are distinct from those in homogeneous solutions and fit for synaptic functions. Thus, we have built a molecular platform for understanding how neuronal synapses are formed and dynamically regulated.

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Section: Psd Condensates Formation By Mixing Major Psd Scaffold Proteinssupporting

confidence: 80%

Section: Methods Details Protein Expression and Purificationmentioning

confidence: 99%

Reconstituted Postsynaptic Density as a Molecular Platform for Understanding Synapse Formation and Plasticity

Zeng

Chen

Guan

et al. 2018

Cell

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354

407

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“…In 320 order to gain insight into the nature of the binding of Gnb5 to the PSD-95 GK domain, we 321 compared PSD-95 -Gnb5 binding to PSD-95 -GKAP binding. GKAP ('GK'-associated protein, 322 also referred to as SAPAP1 or DLGAP1) is an established synaptic GK domain binder (Kim et 323 al., 1997) whose interaction involves the GMP-binding region (Zhu et al, 2017). These ideas 324 are also validated by our own coimmunoprecipitation experiments: GKAP can be efficiently 325 coprecipitated upon pull-down of either the isolated GK domain or an intact PSG module, 326…”

Section: Psd-95 Interactors Occupy Different Gk Subdomains 313mentioning

confidence: 75%

Intramolecular domain dynamics regulate synaptic MAGUK protein interactions

Rademacher

Kuropka²,

Kunde

et al. 2018

Preprint

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21PSD-95 MAGUK family scaffold proteins are multi-domain organisers of synaptic transmission 22 that contain three PDZ domains followed by an SH3-GK domain tandem. This domain 23 architecture allows coordinated assembly of protein complexes composed of neurotransmitter 24 receptors, synaptic adhesion molecules, cytoskeletal proteins and downstream signalling 25 effectors. Here we show that binding of monomeric PDZ3 ligands to the third PDZ domain of 26 PSD-95 induces functional changes in the intramolecular SH3-GK domain assembly that 27 influence subsequent homotypic and heterotypic complex formation. We identify PSD-95 28 interactors that differentially bind to the SH3-GK domain tandem depending on its 29 conformational state. Among these interactors we further establish the heterotrimeric G protein 30 subunit Gnb5 as a PSD-95 complex partner at dendritic spines. The PSD-95 GK domain binds 31 to Gnb5 and this interaction is triggered by PDZ3 ligands binding to the third PDZ domain of 32 PSD-95, unraveling a hierarchical binding mechanism of PSD-95 complex formation. 33 2005). The MAGUK SH3 domain lost its function to bind proline-rich peptides; instead it forms 50 an intramolecular interaction with the GK domain (McGee et al., 2001). Similarly, the PSD-95 51 GK domain is atypical in that it is unable to phosphorylate GMP but has evolved as a protein 52 interaction domain (Johnston et al., 2011). Until now, binding of known interactors to the 53 GK domain typically involves residues of the canonical GMP-binding region (Reese et al., 54 2007; Zhu et al., 2011; Zhu et al., 2016b). This modular array of protein interaction domains 55 allows PSD-95 MAGUKs to function as bidirectional organisers of synaptic function. First, 56 neurotransmitter receptors can be incorporated or removed from postsynaptic membranes, 57 depending on molecular interactions with these sub-membrane scaffold proteins. Second, 58 together with other scaffold proteins at postsynaptic sites, they align downstream effectors and 59 cytoskeletal proteins. Accordingly, PSD-95 family MAGUKs are essential for the establishment 60 of long-term potentiation (LTP) by regulating the content of AMPA receptors at dendritic spines 61 (Ehrlich and Malinow, 2004; Opazo et al., 2012; Sheng et al., 2018). In line with this is the 62 observation that acute knockdown of PSD-95 MAGUKs leads to a decrease in postsynaptic 63 AMPA and NMDA receptor-mediated synaptic transmission as well as a reduction in PSD size 64 (Chen et al., 2015). Taken together, exploring protein complex formation directed by PSD-95 65 MAGUK family members is of central interest to understand synaptic regulation. We have 66 previously shown that synaptic MAGUK proteins oligomerise upon binding of monomeric 67 PDZ3 ligands (ligands that specifically bind to the third PDZ domain) (Rademacher et al., 2013) 68and speculated that ligand -PDZ3 domain binding induces conformational changes in the 69 C-terminal domains that lead to complex formation. Our initial observations of 70 (DTKNYKQASA, referred...

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“…Parallel, a series of clinical imaging studies suggest abnormal synaptic pruning patterns in, among others, frontostriatal brain structures as a possible explanation for volumetric abnormalities in paediatric OCD patients [58,59]. On the other hand, the lower [ 11 C] UCB-J availability in 3-mo old Sapap3 ko mice could also be explained by deviations in synaptogenesis secondary to disruption of the PSD-95/Sapap/Shank complex, with its pivotal role in synaptic development and function [21]. Like Sapap3 ko mice, both PSD-95 ko mice and multiple Shank mutant mouse models show excessive self-grooming and anxiety-like behaviour [51,52].…”

Section: Discussionmentioning

confidence: 99%

“…The spatial and the temporal organization of these components plays a central role in synaptic signalling and plasticity [20]. A key aggregate in this structure is the PSD-95/Sapap/Shank complex, which is crucial for synaptic development and transmission [21]. Mice lacking Sapap3 display synaptic dysfunction with aberrant neurotransmission at the level of (cortico)striatal synapses [11,17,[22][23][24][25][26], resulting in a phenotype suggestive for OCD with excessive grooming and increased anxiety-like behaviour.…”

Section: Introductionmentioning

confidence: 99%

Sapap3 deletion causes dynamic synaptic density abnormalities: a longitudinal [11C]UCB-J PET study in a model of obsessive–compulsive disorder-like behaviour

et al. 2020

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Background Currently, the evidence on synaptic abnormalities in neuropsychiatric disorders—including obsessive–compulsive disorder (OCD)—is emerging. The newly established positron emission tomography (PET) ligand ((R)-1-((3-((11)C-methyl-(11)C)pyridin-4-yl)methyl)-4-(3,4,5-trifluorophenyl)pyrrolidin-2-one) ([11C]UCB-J) provides the opportunity to visualize synaptic density changes in vivo, by targeting the synaptic vesicle protein 2A (SV2A). Here, we aim to evaluate such alterations in the brain of the SAP90/PSD-95-associated protein 3 (Sapap3) knockout (ko) mouse model, showing an abnormal corticostriatal neurotransmission resulting in OCD-like behaviour. Methods Longitudinal [11C]UCB-J µPET/CT scans were acquired in Sapap3 ko and wildtype (wt) control mice (n = 9/group) to study SV2A availability. Based on the Logan reference method, we calculated the volume of distribution (VT(IDIF)) for [11C]UCB-J. Both cross-sectional (wt vs. ko) and longitudinal (3 vs. 9 months) volume-of-interest-based statistical analysis and voxel-based statistical parametric mapping were performed. Both [11C]UCB-J ex vivo autoradiography and [3H]UCB-J in vitro autoradiography were used for the validation of the µPET data. Results At the age of 3 months, Sapap3 ko mice are already characterized by a significantly lower SV2A availability compared to wt littermates (i.a. cortex − 12.69%, p < 0.01; striatum − 14.12%, p < 0.001, thalamus − 13.11%, p < 0.001, and hippocampus − 12.99%, p < 0.001). Healthy ageing in control mice was associated with a diffuse and significant (p < 0.001) decline throughout the brain, whereas in Sapap3 ko mice this decline was more confined to the corticostriatal level. A strong linear relationship (p < 0.0001) was established between the outcome parameters of [11C]UCB-J µPET and [11C]UCB-J ex vivo autoradiography, while such relationship was absent for [3H]UCB-J in vitro autoradiography. Conclusions [11C]UCB-J PET is a potential marker for synaptic density deficits in the Sapap3 ko mouse model for OCD, parallel to disease progression. Our data suggest that [11C]UCB-J ex vivo autoradiography is a suitable proxy for [11C]UCB-J PET data in mice.

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Synaptic Targeting and Function of SAPAPs Mediated by Phosphorylation-Dependent Binding to PSD-95 MAGUKs

Cited by 52 publications

References 67 publications

Reconstituted Postsynaptic Density as a Molecular Platform for Understanding Synapse Formation and Plasticity

Reconstituted Postsynaptic Density as a Molecular Platform for Understanding Synapse Formation and Plasticity

Intramolecular domain dynamics regulate synaptic MAGUK protein interactions

Sapap3 deletion causes dynamic synaptic density abnormalities: a longitudinal [11C]UCB-J PET study in a model of obsessive–compulsive disorder-like behaviour

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