Complement component 5a receptor oligomerization and homologous receptor down-regulation.
Most G-protein-coupled receptors (GPCRs) form di(oligo)-meric structures that constitute signaling and trafficking units and might be essential for receptor functions. Cell responses to complement C5a receptor (C5aR) are tightly controlled by receptor desensitization and internalization. To examine the implication of dimerization in C5aR regulation, we generated an NH(2)-terminally modified C5aR mutant, unable to bind C5a, and a phosphorylation-deficient mutant. Neither an intact NH(2) terminus nor the presence of COOH-terminal phosphorylation sites appeared to be required for the formation of C5aR dimers. Upon C5a stimulation, mutant receptors did not internalize when individually expressed. C5a stimulation of cells that co-expressed wild type C5aR together with either unliganded or phosphorylation-deficient mutant resulted in co-internalization of mutant receptors with C5aR. Unliganded GPCRs can be cross-phosphorylated within a heterologous receptor dimer or by second messenger-activated kinases. C5a stimulation of (32)P-labeled cells that co-expressed the unliganded mutant with either C5aR or the phosphorylation-deficient mutant did not induce phosphorylation of the unliganded mutant. We can thus postulate that, in the case of C5aR, the stimulation and phosphorylation of one monomer is enough to lead to dimer internalization. The existence and functional implication of di(oligo)mer formation may be important for an accurate C5aR down-regulation in pathological conditions.
Rabiet
Marie-Josèphe
MJ
Laboratoire Biochimie et Biophysique des Systèmes Intégrés, Grenoble F-38054, France. marie-josephe.rabiet@cea.fr
Huet
Emilie
E
Boulay
François
F
D002460:Cell Line; D015936:Complement C5a / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D015536:Down-Regulation / Q000502:physiology*; D006801:Humans; D009154:Mutation; D010766:Phosphorylation / Q000235:genetics; D011940:Receptor Aggregation / Q000502:physiology*; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*
C488033:C5AR1 protein, human; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement; D043562:Receptors, G-Protein-Coupled; D015936:Complement C5a
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