PUBLICATION

Cloning and activation of the bullfrog apelin receptor: Gi/o coupling and high affinity for [Pro1]apelin-13

Authors
Moon, M.J., Oh, D.Y., Moon, J.S., Kim, D.K., Hwang, J.I., Lee, J.Y., Kim, J.I., Cho, S., Kwon, H.B., and Seong, J.Y.
ID
ZDB-PUB-090210-20
Date
2007
Source
Molecular and Cellular Endocrinology   277(1-2): 51-60 (Journal)
Registered Authors
Keywords
APJ, Apelin, Bullfrog, Gi/o, GPCR
MeSH Terms
  • Gene Expression Regulation
  • Male
  • Sequence Alignment
  • Molecular Sequence Data
  • Female
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • Humans
  • Signal Transduction
  • Amino Acid Sequence
  • Animals
  • Proline/metabolism*
  • Gene Expression Profiling
  • Cloning, Molecular
  • Ligands
  • GTP-Binding Protein alpha Subunits, Gi-Go/metabolism*
  • Base Sequence
  • Rana catesbeiana/genetics*
  • DNA, Complementary/genetics
  • Receptors, G-Protein-Coupled/chemistry
  • Receptors, G-Protein-Coupled/genetics*
  • Receptors, G-Protein-Coupled/metabolism*
  • Cell Line
  • Sequence Homology, Amino Acid
  • Intercellular Signaling Peptides and Proteins/metabolism*
PubMed
17825479 Full text @ Mol. Cell. Endocrinol.
Abstract
In mammals, apelin and its G protein-coupled receptor, APJ, regulate blood pressure, intake of food and water, and cardiac contractility. In this study, we report the cloning and functional characterization of APJ in the bullfrog, Rana catesbeiana. Bullfrog APJ (bfAPJ) cDNA contains an open reading frame of 1083 nucleotides encoding a protein of 360 amino acid residues. Sequence alignment reveals 75% amino acid identity with Xenopus, 63% identity with zebrafish and 40-42% identity with mammalian APJs. RT-PCR analysis and tissue binding assay reveal high expression of bfAPJ mRNA in the brain, particularly in the hypothalamus, and moderate expression in the pituitary, testis, adrenal gland and lung. Whereas [pGlu(1)]apelin-13 did not induce CRE-luc (protein kinase A-specific reporter) and SRE-luc (protein kinase C-specific reporter) activity in cells expressing bfAPJ, this apelin-13 decreased forskolin-induced CRE-luc activity and cAMP accumulation in a pertussis toxin-sensitive manner. This study indicates that bfAPJ may couple to G(i/o). [Pro(1)]apelin-13, a synthetic apelin based on the sequence of the putative apelin gene from many non-mammalian species, activates bfAPJ with 5-10-fold greater sensitivity/affinity than mammalian apelin-13. Collectively, this study expands our understanding of the physiological roles of this receptor system in non-mammalian species.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping