PUBLICATION
Unexpected role of a conserved domain in the first extracellular loop in G protein-coupled receptor trafficking
- Authors
- Rizzo, M.J., Evans, J.P., Burt, M., Saunders, C.J., Johnson, E.C.
- ID
- ZDB-PUB-190402-20
- Date
- 2018
- Source
- Biochemical and Biophysical Research Communications 503: 1919-1926 (Journal)
- Registered Authors
- Johnson, Eric
- Keywords
- G protein-coupled receptor (GPCR), Membrane trafficking, Membrane transport, Mutagenesis, Signaling
- MeSH Terms
-
- Animals
- Caenorhabditis elegans
- Chickens
- Ciona intestinalis
- Cloning, Molecular
- Drosophila melanogaster
- Humans
- Models, Molecular
- Mutagenesis, Site-Directed
- Receptors, G-Protein-Coupled/chemistry
- Receptors, G-Protein-Coupled/genetics
- Receptors, G-Protein-Coupled/metabolism*
- Signal Transduction/genetics
- Xenopus laevis
- Zebrafish
- PubMed
- 30064912 Full text @ Biochem. Biophys. Res. Commun.
Citation
Rizzo, M.J., Evans, J.P., Burt, M., Saunders, C.J., Johnson, E.C. (2018) Unexpected role of a conserved domain in the first extracellular loop in G protein-coupled receptor trafficking. Biochemical and Biophysical Research Communications. 503:1919-1926.
Abstract
G protein-coupled receptors are the largest superfamily of cell surface receptors in the Metazoa and play critical roles in transducing extracellular signals into intracellular responses. This action is mediated through conformational changes in the receptor following ligand binding. A number of conserved motifs have critical roles in GPCR function, and here we focus on a highly conserved motif (WxFG) in extracellular loop one (EL1). A phylogenetic analysis documents the presence of the WxFG motif in ∼90% of Class A GPCRs and the motif is represented in 17 of the 19 Class A GPCR subfamilies. Using site-directed mutagenesis, we mutagenized the conserved tryptophan residue in eight receptors which are members of disparate class A GPCR subfamilies from different taxa. The modification of the Drosophila leucokinin receptor shows that substitution of any non-aromatic amino acid for the tryptophan leads to a loss of receptor function. Additionally, leucine substitutions at this position caused similar signaling defects in the follicle-stimulating hormone receptor (FSHR), Galanin receptor (GALR1), AKH receptor (AKHR), corazonin receptor (CRZR), and muscarinic acetylcholine receptor (mACHR1). Visualization of modified receptors through the incorporation of a fluorescent tag revealed a severe reduction in plasma membrane expression, indicating aberrant trafficking of these modified receptors. Taken together, these results suggest a novel role for the WxFG motif in GPCR trafficking and receptor function.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping