Molecular cloning and functional expression of the first insect FMRFamide receptor.
FMRFamide and FMRFamide-related neuropeptides are extremely widespread and abundant in invertebrates and have numerous important functions. Here, we have cloned a Drosophila orphan receptor, and stably expressed it in Chinese hamster ovary cells. Screening of a peptide library revealed that the receptor reacted with high affinity to FMRFamide (EC50, 6 x 10(-9) M). The intrinsic Drosophila FMRFamide peptides are known to be synthesized as a large preprohormone, containing at least 13 related FMRFamide peptides (8 distinct FMRFamides). Screening of these intrinsic Drosophila FMRFamides showed that the receptor had highest affinity to Drosophila FMRFamide-6 (PDNFMRFamide) (EC50, 9 x 10(-10) M), whereas it had a somewhat lower affinity to Drosophila FMRFamide-2 (DPKQDFMRFamide) (EC50, 3 x 10(-9) M) and considerably less affinity to the other Drosophila FMRFamide-related peptides. To our knowledge, this article is the first report on the molecular identification of an invertebrate FMRFamide receptor.
Cazzamali
Giuseppe
G
Department of Cell Biology, Zoological Institute, University of Copenhagen, Universitetsparken 15, DK-2100 Copenhagen, Denmark.
Grimmelikhuijzen
Cornelis J P
CJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism*; D019835:FMRFamide / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D017344:Genes, Insect; D008969:Molecular Sequence Data; D019151:Peptide Library; D018010:Receptors, Invertebrate Peptide / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; C055209:FMRFamide receptor; D019151:Peptide Library; D018010:Receptors, Invertebrate Peptide; D011994:Recombinant Proteins; D019835:FMRFamide