Identification of one capa and two pyrokinin receptors from the malaria mosquito Anopheles gambiae.

Abstract

We cloned the cDNA of three evolutionarily related G protein-coupled receptors from the malaria mosquito Anopheles gambiae and functionally expressed them in Chinese hamster ovary cells. One receptor, Ang-Capa-R, was only activated by the two Anopheles capa neuropeptides Ang-capa-1 (GPTVGLFAFPRVamide) and Ang-capa-2 (pQGLVPFPRVamide) with EC(50) values of 8.6x10(-9)M and 3.3x10(-9)M, respectively, but not by any other known mosquito neuropeptide. The second receptor, Ang-PK-1-R, was selectively activated by the Anopheles pyrokinin-1 peptides Ang-PK-1-1 (AGGTGANSAMWFGPRLamide) and Ang-PK-1-2 (AAAMWFGPRLamide) with EC(50) values of 3.3x10(-8)M and 2.5x10(-8)M, respectively, but not by mosquito capa or pyrokinin-2 peptides. For the third receptor, Ang-PK-2-R, the most potent ligands were the pyrokinin-2 peptides Ang-PK-2-1 (DSVGENHQRPPFAPRLamide) and Ang-PK-2-2 (NLPFSPRLamide) with EC(50) values of 5.2x10(-9)M and 6.4x10(-9)M, respectively. However, this receptor could also be activated by the two pyrokinins-1, albeit with lower potency (EC(50): 2-5x10(-8)M). Because Ang-capa-1 and -2 and Ang-PK-1-1 are located on one preprohormone and the other peptides on another prohormone, these results imply a considerable crosstalk between the capa, pyrokinin-1 and pyrokinin-2 systems. Gene structure and phylogenetic tree analyses showed that Ang-Capa-R was the orthologue of the Drosophila capa receptor CG14575, Ang-PK-1-R the orthologue of the Drosophila pyrokinin-1 receptor CG9918, and Ang-PK-2-R the orthologue of the Drosophila pyrokinin-2 receptors CG8784 and CG8795. This is the first report on the functional characterization and crosstalk properties of capa and pyrokinin receptors in mosquitoes.

• Authors: Stine S Olsen, Giuseppe Cazzamali, Michael Williamson, Cornelis J P Grimmelikhuijzen, Frank Hauser
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 362 Issue 2 Pg. 245-51 (Oct 19 2007) ISSN: 0006-291X [Print] United States
• PMID: 17709098 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA, Complementary
• Insect Proteins
• Neuropeptides
• Receptors, G-Protein-Coupled
• Receptors, Neuropeptide
• pyrokinin
• Green Fluorescent Proteins
• GTP-Binding Proteins

Topics

• Amino Acid Sequence
• Animals
• Anopheles (genetics, metabolism)
• CHO Cells
• Cricetinae
• Cricetulus
• DNA, Complementary (genetics, isolation & purification)
• GTP-Binding Proteins (genetics, metabolism)
• Green Fluorescent Proteins (genetics, metabolism)
• Humans
• Insect Proteins (genetics, physiology)
• Insect Vectors (genetics, metabolism)
• Malaria (parasitology)
• Molecular Sequence Data
• Neuropeptides (genetics, metabolism, pharmacology)
• Phylogeny
• Receptor Cross-Talk (drug effects, physiology)
• Receptors, G-Protein-Coupled (agonists, genetics, physiology)
• Receptors, Neuropeptide (agonists, genetics, physiology)
• Sequence Alignment
• Sequence Homology, Amino Acid
• Transfection