DOI: 10.1101/462358Nov 5, 2018Paper

Intramolecular domain dynamics regulate synaptic MAGUK protein interactions

BioRxiv : the Preprint Server for Biology
Nils RademacherSarah A Shoichet

Abstract

PSD-95 MAGUK family scaffold proteins are multi-domain organisers of synaptic transmission that contain three PDZ domains followed by an SH3-GK domain tandem. This domain architecture allows coordinated assembly of protein complexes composed of neurotransmitter receptors, synaptic adhesion molecules, cytoskeletal proteins and downstream signalling effectors. Here we show that binding of monomeric PDZ3 ligands to the third PDZ domain of PSD-95 induces functional changes in the intramolecular SH3-GK domain assembly that influence subsequent homotypic and heterotypic complex formation. We identify PSD-95 interactors that differentially bind to the SH3-GK domain tandem depending on its conformational state. Among these interactors we further establish the heterotrimeric G protein subunit Gnb5 as a PSD-95 complex partner at dendritic spines. The PSD-95 GK domain binds to Gnb5 and this interaction is triggered by PDZ3 ligands binding to the third PDZ domain of PSD-95, unraveling a hierarchical binding mechanism of PSD-95 complex formation.

Methods Mentioned

BETA
acetylation
pull-down
FCS
flow cytometry
transfection
PCR
CoIP
coimmunoprecipitation
pull-downs
confocal microscopy

Software Mentioned

Mascot Distiller Quantitation Toolbox

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