Synaptic MAGUK multimer formation is mediated by PDZ domains and promoted by ligand binding.
Chem Biol
; 20(8): 1044-54, 2013 Aug 22.
Article
in En
| MEDLINE
| ID: mdl-23973190
ABSTRACT
To examine the scaffolding properties of PSD-95, we have taken advantage of established ligand/PDZ domain interactions and developed a cell-based assay for investigating protein complex formation. This assay enables quantitative analysis of PDZ domain-mediated protein clustering using bimolecular fluorescence complementation (BiFC). Two nonfluorescent halves of EYFP were fused to C-terminal PDZ ligand sequences to generate probes that sense for PDZ domain binding grooves of adjacent (interacting) molecules. When these probes are brought into proximity by the PDZ domains of a multiprotein scaffold, a functional fluorescent EYFP molecule can be detected. We have used this system to examine the properties of selected PSD-95 variants and thereby delineated regions of importance for PSD-95 complex formation. Further analysis led to the finding that PSD-95 multimerization is PDZ domain-mediated and promoted by ligand binding.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Synapses
/
Guanylate Kinases
/
PDZ Domains
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Chem Biol
Journal subject:
BIOLOGIA
/
BIOQUIMICA
/
QUIMICA
Year:
2013
Document type:
Article
Affiliation country:
Germany