ZFIN ID: ZDB-PUB-020618-3
Cloning, structural characterization and functional expression of a zebrafish bradykinin B2-related receptor
Dunér, T., Conlon, J.M., Kukkonen, J.P., Akerman, K.E.O., Yan, Y.-L., Postlethwait, J.H., and Larhammar, D.
Date: 2002
Source: The Biochemical journal 364(3): 817-824 (Journal)
Registered Authors: Dunér, Torun, Larhammar, Dan, Postlethwait, John H., Yan, Yi-Lin
Keywords: Ca2+; chromosome; G-protein-coupled receptor; evolution; teleost; trout
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Cell Line
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA Primers
  • Gene Expression Regulation
  • Genetic Vectors
  • Humans
  • Molecular Sequence Data
  • Phylogeny
  • Polymerase Chain Reaction
  • Receptor, Bradykinin B2
  • Receptors, Bradykinin/chemistry
  • Receptors, Bradykinin/genetics*
  • Receptors, Bradykinin/metabolism
  • Recombinant Proteins/metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Transfection
  • Zebrafish/genetics*
PubMed: 12049646 Full text @ Biochem. J.
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ABSTRACT
The actions of bradykinin (BK) in mammals are mediated through the activation of the B1 and B2 BK receptors. The only BK receptor that has been cloned from a non-mammalian species is a B2-like receptor from the chicken (termed the ornithokinin receptor). Pharmacological studies have demonstrated the presence of BK receptors in tissues of teleost fishes , such as trout and cod, but the ligand-binding properties of these receptors differ appreciably from those of the mammalian and chicken receptors. We report here the cloning of a B2-like receptor in zebrafish that shares 35% identity with human B2 and 30% identity with human B1. Phylogenetic analyses confirm a closer relationship with B2 than B1. The receptor gene was mapped to linkage group 17, which is syntenic to the human B2-B1 gene region. After functional expression of the zebrafish B2 receptor in mammalian cells, nanomolar concentrations of trout BK ([Arg (0),Trp(5),Leu(8)]-BK) and the derivative [des-Arg(0),Trp(5),Leu(8)]-BK (where 'des' indicates a missing amino acid) induced a significant transient rise in intracellular free Ca(2+). The B1-selective analogue [ Arg(0),Trp(5),Leu(8),des-Arg(9)]-BK was inactive at nanomolar concentrations. Taken together, these results strongly support the gene' s identity as a piscine orthologue of the mammalian B2 receptor.
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