Mammalian calcitonin receptor-like receptor/receptor activity modifying protein complexes define calcitonin gene-related peptide and adrenomedullin receptors in Drosophila Schneider 2 cells

FEBS Lett. 2000 Apr 14;471(2-3):156-60. doi: 10.1016/s0014-5793(00)01387-9.

Abstract

Differential glycosylation of human and rat (r) calcitonin (CT) receptor-like receptors (CRLR) as a result of interactions with accessory receptor activity-modifying proteins (RAMP)1 or -2 was considered to define CT gene-related peptide (CGRP) or adrenomedullin (ADM) receptors in mammalian cells. Here, Drosophila Schneider (S2) cells stably co-expressed rCRLR and RAMP1 or -2 as functional CGRP or ADM receptors. Different from mammalian cells, rCRLR expressed in S2 cells are uniformly glycosylated proteins independent of RAMP1 or RAMP2. Bis(sulfosuccinimidyl)suberate cross-linking revealed receptor components with the size of rCRLR, increased by the molecular weights of the corresponding RAMPs and [(125)I]CGRP or [(125)I]ADM. In conclusion, [(125)I]CGRP/rCRLR/RAMP1 and [(125)I]ADM/rCRLR/RAMP2 complexes have been recognized in Drosophila S2 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenomedullin
  • Amidohydrolases / metabolism
  • Animals
  • Blotting, Western
  • Calcitonin Gene-Related Peptide / antagonists & inhibitors
  • Calcitonin Gene-Related Peptide / chemistry
  • Calcitonin Gene-Related Peptide / metabolism*
  • Calcitonin Receptor-Like Protein
  • Cell Line
  • Cross-Linking Reagents / metabolism
  • Cyclic AMP / metabolism
  • Drosophila melanogaster / cytology*
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / metabolism
  • Glycosylation
  • Humans
  • Inhibitory Concentration 50
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Molecular Weight
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
  • Peptides / antagonists & inhibitors
  • Peptides / chemistry
  • Peptides / metabolism
  • Protein Binding
  • Rats
  • Receptor Activity-Modifying Protein 1
  • Receptor Activity-Modifying Protein 2
  • Receptor Activity-Modifying Proteins
  • Receptors, Adrenomedullin
  • Receptors, Calcitonin / chemistry
  • Receptors, Calcitonin / genetics
  • Receptors, Calcitonin / metabolism*
  • Receptors, Cell Surface / chemistry
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism
  • Receptors, Peptide / antagonists & inhibitors
  • Receptors, Peptide / chemistry
  • Receptors, Peptide / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Succinimides / metabolism
  • Transfection

Substances

  • CALCRL protein, human
  • Calcitonin Receptor-Like Protein
  • Calcrl protein, rat
  • Cross-Linking Reagents
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Peptides
  • RAMP1 protein, human
  • RAMP2 protein, human
  • Ramp1 protein, rat
  • Ramp2 protein, rat
  • Receptor Activity-Modifying Protein 1
  • Receptor Activity-Modifying Protein 2
  • Receptor Activity-Modifying Proteins
  • Receptors, Adrenomedullin
  • Receptors, Calcitonin
  • Receptors, Cell Surface
  • Receptors, Peptide
  • Recombinant Fusion Proteins
  • Succinimides
  • Adrenomedullin
  • Cyclic AMP
  • bis(sulfosuccinimidyl)suberate
  • Amidohydrolases
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
  • Calcitonin Gene-Related Peptide