Reverse physiology in drosophila: identification of a novel allatostatin-like neuropeptide and its cognate receptor structurally related to the mammalian somatostatin/galanin/opioid receptor family

EMBO J. 1999 Nov 1;18(21):5892-900. doi: 10.1093/emboj/18.21.5892.

Abstract

By using degenerate oligonucleotide primers deduced from the conserved regions of the mammalian somatostatin receptors, a novel G-protein-coupled receptor from Drosophila melanogaster has been isolated exhibiting structural similarities to mammalian somatostatin/galanin/opioid receptors. To identify the bioactive ligand, a 'reverse physiology' strategy was used whereby orphan Drosophila receptor-expressing frog oocytes were screened against potential ligands. Agonistic activity was electrophysiologically recorded as inward potassium currents mediated through co-expressed G-protein-gated inwardly rectifying potassium channels (GIRK). Using this approach a novel peptide was purified from Drosophila head extracts. Mass spectrometry revealed an octapeptide of 925 Da with a sequence Ser-Arg-Pro-Tyr-Ser-Phe-Gly-Leu-NH(2) reminiscent of insect allatostatin peptides known to control diverse functions such as juvenile hormone synthesis during metamorphosis or visceral muscle contractions. Picomolar concentrations of the synthesized octapeptide activated the cognate receptor response mediated through GIRK1, indicating that we have isolated the 394-amino-acid Drosophila allatostatin receptor which is coupled to the Gi/Go class of G proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • Drosophila Proteins*
  • Drosophila melanogaster / chemistry*
  • Evolution, Molecular
  • GTP-Binding Proteins / chemistry
  • Gene Expression
  • Insect Proteins*
  • Mass Spectrometry
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Molecular Sequence Data
  • Neuropeptides / chemistry*
  • Neuropeptides / genetics*
  • Oocytes
  • RNA, Messenger / metabolism
  • Receptors, Cell Surface / chemistry*
  • Receptors, G-Protein-Coupled*
  • Receptors, Galanin
  • Receptors, Neuropeptide / chemistry
  • Receptors, Neuropeptide / genetics*
  • Receptors, Opioid / chemistry
  • Receptors, Somatostatin / chemistry
  • Sequence Homology, Amino Acid
  • Xenopus

Substances

  • AstA-R1 protein, Drosophila
  • Drosophila Proteins
  • Insect Proteins
  • Membrane Proteins
  • Neuropeptides
  • RNA, Messenger
  • Receptors, Cell Surface
  • Receptors, G-Protein-Coupled
  • Receptors, Galanin
  • Receptors, Neuropeptide
  • Receptors, Opioid
  • Receptors, Somatostatin
  • allatostatin
  • GTP-Binding Proteins

Associated data

  • GENBANK/AF163775