title: Substitution of the cysteine 438 residue in the cytoplasmic tail of the glucagon-like peptide-1 receptor alters signal transduction activity.
abstract: Several G-protein-coupled receptors contain cysteine residues in the C-terminal tail that may modulate receptor function. In this work we analysed the substitution of Cys438 by alanine in the glucagon-like peptide-1 (GLP-1) receptor (GLPR), which led to a threefold decrease in cAMP production, although endocytosis and cellular redistribution of GLP-1 receptor agonist-induced processes were unaffected. Additionally, cysteine residues in the C-terminal tail of several G-protein-coupled receptors were found to act as substrates for palmitoylation, which might modify the access of protein kinases to this region. His-tagged GLP-1 receptors incorporated 3H-palmitate. Nevertheless, substitution of Cys438 prevented the incorporation of palmitate. Accordingly, we also investigated the effect of substitution of the consensus sequence by protein kinase C (PKC) Ser431/432 in both wild-type and Ala438 GLP-1 receptors. Substitution of Ser431/432 by alanine did not modify the ability of wild-type receptors to stimulate adenylate cyclase or endocytosis and recycling processes. By contrast, the substitution of Ser431/432 by alanine in the receptor containing Ala438 increased the ability to stimulate adenylate cyclase. All types of receptors were mainly internalised through coated pits. Thus, cysteine 438 in the cytoplasmic tail of the GLP-1 receptor would regulate its interaction with G-proteins and the stimulation of adenylyl cyclase. Palmitoylation of this residue might control the access of PKC to Ser431/432.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Complutense de Madrid, Ciudad Universitaria, 28040 Madrid, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Vázquez
authors.forename: Isabel
authors.initials: I
authors.lastname: Roncero
authors.forename: Enrique
authors.initials: E
authors.lastname: Blázquez
authors.forename: Elvira
authors.initials: E
authors.lastname: Alvarez
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000409:Alanine; D000818:Animals; D016466:CHO Cells; D016384:Consensus Sequence; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D003545:Cysteine / Q000235:genetics*; D003593:Cytoplasm / Q000378:metabolism*; D004705:Endocytosis; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism; D000067757:Glucagon-Like Peptide-1 Receptor; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010168:Palmitates / Q000378:metabolism; D011493:Protein Kinase C / Q000235:genetics; D051381:Rats; D018027:Receptors, Glucagon / Q000235:genetics*; D012694:Serine; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection
chemical_list: C000593835:Glp1r protein, rat; D000067757:Glucagon-Like Peptide-1 Receptor; D010168:Palmitates; D018027:Receptors, Glucagon; D012694:Serine; D000242:Cyclic AMP; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D003545:Cysteine; D000409:Alanine