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
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Research Support, Non-U.S. Gov't
MeSH terms
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Adrenomedullin
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Amidohydrolases / metabolism
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Animals
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Blotting, Western
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Calcitonin Gene-Related Peptide / antagonists & inhibitors
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Calcitonin Gene-Related Peptide / chemistry
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Calcitonin Gene-Related Peptide / metabolism*
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Calcitonin Receptor-Like Protein
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Cell Line
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Cross-Linking Reagents / metabolism
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Cyclic AMP / metabolism
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Drosophila melanogaster / cytology*
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Drosophila melanogaster / genetics
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Drosophila melanogaster / metabolism
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Glycosylation
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Humans
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Inhibitory Concentration 50
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Intracellular Signaling Peptides and Proteins
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Molecular Weight
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Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
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Peptides / antagonists & inhibitors
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Peptides / chemistry
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Peptides / metabolism
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Protein Binding
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Rats
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Receptor Activity-Modifying Protein 1
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Receptor Activity-Modifying Protein 2
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Receptor Activity-Modifying Proteins
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Receptors, Adrenomedullin
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Receptors, Calcitonin / chemistry
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Receptors, Calcitonin / genetics
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Receptors, Calcitonin / metabolism*
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Receptors, Cell Surface / chemistry
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Receptors, Cell Surface / genetics
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Receptors, Cell Surface / metabolism
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Receptors, Peptide / antagonists & inhibitors
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Receptors, Peptide / chemistry
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Receptors, Peptide / metabolism*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Succinimides / metabolism
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Transfection
Substances
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CALCRL protein, human
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Calcitonin Receptor-Like Protein
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Calcrl protein, rat
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Cross-Linking Reagents
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Intracellular Signaling Peptides and Proteins
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Membrane Proteins
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Peptides
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RAMP1 protein, human
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RAMP2 protein, human
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Ramp1 protein, rat
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Ramp2 protein, rat
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Receptor Activity-Modifying Protein 1
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Receptor Activity-Modifying Protein 2
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Receptor Activity-Modifying Proteins
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Receptors, Adrenomedullin
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Receptors, Calcitonin
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Receptors, Cell Surface
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Receptors, Peptide
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Recombinant Fusion Proteins
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Succinimides
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Adrenomedullin
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Cyclic AMP
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bis(sulfosuccinimidyl)suberate
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Amidohydrolases
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Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
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Calcitonin Gene-Related Peptide