Olsen Stine S, Cazzamali Giuseppe, Williamson Michael, Grimmelikhuijzen Cornelis J P, Hauser Frank
Center for Functional and Comparative Insect Genomics, Department of Cell Biology and Comparative Zoology, Institute of Biology, University of Copenhagen, Universitetsparken 15, DK-2100 Copenhagen, Denmark.
Biochem Biophys Res Commun. 2007 Oct 19;362(2):245-51. doi: 10.1016/j.bbrc.2007.06.190. Epub 2007 Jul 27.
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.
我们从冈比亚疟蚊中克隆了三种进化相关的G蛋白偶联受体的cDNA，并在中国仓鼠卵巢细胞中对其进行了功能表达。其中一种受体，即Ang-Capa-R，仅被两种冈比亚疟蚊CAPA神经肽Ang-capa-1（GPTVGLFAFPRVamide）和Ang-capa-2（pQGLVPFPRVamide）激活，其半数有效浓度（EC50）值分别为8.6×10−9M和3.3×10−9M，而不被任何其他已知的蚊虫神经肽激活。第二种受体，即Ang-PK-1-R，被疟蚊速激肽-1肽Ang-PK-1-1（AGGTGANSAMWFGPRLamide）和Ang-PK-1-2（AAAMWFGPRLamide）选择性激活，其EC50值分别为3.3×10−8M和2.5×10−8M，但不被蚊虫CAPA或速激肽-2肽激活。对于第三种受体，即Ang-PK-2-R，最有效的配体是速激肽-2肽Ang-PK-2-1（DSVGENHQRPPFAPRLamide）和Ang-PK-2-2（NLPFSPRLamide），其EC50值分别为5.2×10−9M和6.4×10−9M。然而，该受体也可被两种速激肽-1激活，尽管效力较低（EC50：2 - 5×10−8M）。由于Ang-capa-1和-2以及Ang-PK-1-1位于一种前激素原上，而其他肽位于另一种前激素原上，这些结果表明CAPA、速激肽-1和速激肽-2系统之间存在相当程度的相互作用。基因结构和系统发育树分析表明，Ang-Capa-R是果蝇CAPA受体CG14575的直系同源物，Ang-PK-1-R是果蝇速激肽-1受体CG9918的直系同源物，Ang-PK-2-R是果蝇速激肽-2受体CG8784和CG8795的直系同源物。这是关于蚊虫中CAPA和速激肽受体的功能特性及相互作用特性的首次报道。
