1 research outputs found
Cell Type-Specific Tandem Affinity Purification of the Mouse Hippocampal CB1 Receptor-Associated Proteome
G protein
coupled receptors (GPCRs) exert their effects through
multiprotein signaling complexes. The cannabinoid receptor type 1
(CB1) is among the most abundant GPCRs in the mammalian brain and
involved in a plethora of physiological functions. We used a combination
of viral-mediated cell type-specific expression of a tagged CB1 fusion
protein (CB1-SF), tandem affinity purification (TAP) and proteomics
on hippocampal mouse tissue to analyze the composition and differences
of CB1 protein complexes in glutamatergic neurons and in GABAergic
interneurons. Purified proteins underwent tryptic digestion and were
identified using deep-coverage data-independent acquisition with ion
mobility separation-enhanced mass spectroscopy, leading to the identification
of 951 proteins specifically enriched in glutamatergic and GABAergic
CB1-SF TAP samples as compared to controls. Gene Ontology and protein
network analyses showed an enrichment of single proteins and functional
clusters of proteins involved in already well described domains of
CB1 functions. Supported by this consistent data set we could confirm
already known CB1 interactors, reveal new potentially interacting
proteins and differences in cell type-specific signaling properties
of CB1, thereby providing the foundation for further functional studies
on differential CB1 signaling