GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.

Have any questions? Please email us at ecoliwiki@gmail.com

PMID:14722252

From GONUTS
Jump to: navigation, search
Citation

Kuwasako, K, Cao, YN, Nagoshi, Y, Tsuruda, T, Kitamura, K and Eto, T (2004) Characterization of the human calcitonin gene-related peptide receptor subtypes associated with receptor activity-modifying proteins. Mol. Pharmacol. 65:207-13

Abstract

Coexpression of receptor activity-modifying proteins (RAMPs) with calcitonin receptor 2 (CTR2) or calcitonin receptor-like receptor (CRLR) leads to the formation of four functional heterodimeric receptors for human calcitonin gene-related peptide (hCGRP). In this study, we transfected hCGRP receptors into human embryonic kidney 293 cells and examined their pharmacological profiles using three dominant-negative (DN) RAMP mutants and various hCGRPalpha analogs. Fluorescence-activated cell-sorting analysis revealed that their cotransfection with CTR2 induced cell surface expression of all three RAMPs, and the three CTR2/RAMP heterodimers mediated equivalent levels of cAMP production in response to hCGRPalpha that were approximately 50-fold greater than were seen with CTR2 alone. By contrast, [Tyr0]hCGRPalpha binding and signaling were markedly weaker with CTR2/RAMP2 or -3 than with CTR2/RAMP1 or CRLR/RAMP1; likewise, 125I-[His10]hCGRPalpha bound most potently to CTR2/RAMP1. When CTR2 was coexpressed with DN RAMP1 or -2, hCGRPalpha-evoked responses were similar to those seen with CTR2 alone, despite the expression of both CTR2 and DN RAMP at the cell surface. But coexpression of DN RAMP3 with CTR2 significantly diminished hCGRPalpha signaling compared with that seen with CTR2 alone, indicating that DN RAMP3 is able to function as a negative regulator of CTR2 function. Competition experiments showed the relative agonist sensitivity of the four receptors to be hCGRPalpha > [Tyr0]hCGRPalpha > [Cys(Et)2,7]hCGRPalpha > [Cys(ACM)2,7]hCGRPalpha. Of the linear analogs, [Cys(ACM)2,7]hCGRPalpha (ACM, acetylmethoxy) enhanced cAMP formation only via CTR2/RAMP1, whereas [Cys(Et2,7)]hCGRPalpha acted via CRLR/RAMP1 and somewhat less potently via CTR2/RAMP1. Thus, among the three CGRP8-37-insensitive receptors, CTR2/RAMP1 is most sensitive to the two linear analogs, suggesting that it could be classified as a CGRP2 receptor. Moreover, the combined use of iodinated CGRPalpha analogs may be useful for defining the CGRP1 receptor.

Links

PubMed Online version:10.1124/mol.65.1.207

Keywords

Cell Line; Cells, Cultured; Humans; Intracellular Signaling Peptides and Proteins; Membrane Proteins/genetics; Membrane Proteins/metabolism; Peptides/pharmacology; Radioligand Assay; Receptor Activity-Modifying Protein 1; Receptor Activity-Modifying Protein 2; Receptor Activity-Modifying Protein 3; Receptor Activity-Modifying Proteins; Receptors, Calcitonin/genetics; Receptors, Calcitonin/metabolism; Receptors, Calcitonin Gene-Related Peptide/genetics; Receptors, Calcitonin Gene-Related Peptide/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:RAMP1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P06881

F

Seeded From UniProt

complete

HUMAN:RAMP1

involved_in

GO:0007189: adenylate cyclase-activating G protein-coupled receptor signaling pathway

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:RAMP1

located_in

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:RAMP1

enables

GO:0001635: calcitonin gene-related peptide receptor activity

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P30988

F

Seeded From UniProt

complete

HUMAN:RAMP1

involved_in

GO:0097647: amylin receptor signaling pathway

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:RAMP1

enables

GO:0097643: amylin receptor activity

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P30988

F

Seeded From UniProt

complete

HUMAN:RAMP2

located_in

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:RAMP3

located_in

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:RAMP3

involved_in

GO:0038041: cross-receptor inhibition within G protein-coupled receptor heterodimer

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P30988

P

  • has_input:(UniProtKB:P30988)

Seeded From UniProt

complete

HUMAN:CALCA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P30988

F

Seeded From UniProt

complete

HUMAN:CALCA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O60894

F

Seeded From UniProt

complete

HUMAN:CALCA

involved_in

GO:0007189: adenylate cyclase-activating G protein-coupled receptor signaling pathway

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CALCA

enables

GO:0044877: protein-containing complex binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:CALCR

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P06881

F

Seeded From UniProt

complete

HUMAN:CALCR

enables

GO:0001635: calcitonin gene-related peptide receptor activity

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O60894

F

Seeded From UniProt

complete

HUMAN:CALCR

involved_in

GO:0007189: adenylate cyclase-activating G protein-coupled receptor signaling pathway

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CALCR

involved_in

GO:0097647: amylin receptor signaling pathway

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CALCR

enables

GO:0097643: amylin receptor activity

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O60894

F

Seeded From UniProt

complete

HUMAN:CALCR

involved_in

GO:0038041: cross-receptor inhibition within G protein-coupled receptor heterodimer

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O60896

P

Seeded From UniProt

complete

See also

References

See Help:References for how to manage references in GONUTS.