Cloning and properties of a novel natriuretic peptide receptor, NPR-D

M. Kashiwagi, T. Katafuchi, A. Kato, H. Inuyama, T. Ito, H. Hagiwara, Y. Takei, S. Hirose

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

A novel natriuretic peptide receptor, which we have termed natriuretic peptide receptor D (NPR-D), has been cloned and characterized. cDNAs related to the natriuretic peptide receptor (NPR) were amplified by PCR from a template of poly(A)-rich RNA isolated from the eel gill. Sequencing of the PCR products revealed the presence of a new clone that showed about 70% sequence identity to the eel type-C receptor, NPR-C. The PCR fragment was used to determine the tissue distribution of the new NPR-D message by an RNase protection assay, which gave the strongest signal in brain samples, and then used to screen a brain library to obtain a full-length cDNA clone. The cDNA clone predicted a protein of 500 amino acids containing a signal sequence and a hydrophobic transmembrane segment. The predicted sequence also contained the NPR motif which is essential for the binding of natriuretic peptides. The protein NPR-D was expressed in COS cells and shown to have high affinities for eel and rat natriuretic peptides. The newly cloned NPR-D has a short cytoplasmic tail; in this respect, NPR-C and NPR-D are very similar and form a subfamily of the NPR family. Affinity labeling indicated that NPR-D exists as a disulfide-linked tetramer. This is a marked contrast to the homodimeric structure of NPR-C, HS-142-1, a non-peptide natriuretic peptide receptor antagonist of microbial origin previously shown to be selective for the guanylate-cyclase-coupled receptors NPR-A and NPR-B, competitively inhibited the binding of 125I-labeled eel natriuretic peptide to eel NPR-D, whereas it did not affect the binding activity of eel NPR-C, suggesting that HS-142-1 is an antagonist that recognizes the tetrameric structures of NPR since the guanylate-cyclase-coupled receptors have also been demonstrated to exist as tetramers.

Original languageEnglish (US)
Pages (from-to)102-109
Number of pages8
JournalEuropean Journal of Biochemistry
Volume233
Issue number1
StatePublished - 1995

Fingerprint

Natriuretic Peptides
Peptide Receptors
Cloning
Organism Cloning
Eels
Guanylate Cyclase-Coupled Receptors
Complementary DNA
Clone Cells
Polymerase Chain Reaction
Brain
Ribonuclease III
C-Type Natriuretic Peptide
Poly A
COS Cells
Ribonucleases
Protein Sorting Signals
Tissue Distribution
Disulfides
Labeling

Keywords

  • antagonist
  • cloning
  • eel
  • natriuretic peptide
  • receptor

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kashiwagi, M., Katafuchi, T., Kato, A., Inuyama, H., Ito, T., Hagiwara, H., ... Hirose, S. (1995). Cloning and properties of a novel natriuretic peptide receptor, NPR-D. European Journal of Biochemistry, 233(1), 102-109.

Cloning and properties of a novel natriuretic peptide receptor, NPR-D. / Kashiwagi, M.; Katafuchi, T.; Kato, A.; Inuyama, H.; Ito, T.; Hagiwara, H.; Takei, Y.; Hirose, S.

In: European Journal of Biochemistry, Vol. 233, No. 1, 1995, p. 102-109.

Research output: Contribution to journalArticle

Kashiwagi, M, Katafuchi, T, Kato, A, Inuyama, H, Ito, T, Hagiwara, H, Takei, Y & Hirose, S 1995, 'Cloning and properties of a novel natriuretic peptide receptor, NPR-D', European Journal of Biochemistry, vol. 233, no. 1, pp. 102-109.
Kashiwagi M, Katafuchi T, Kato A, Inuyama H, Ito T, Hagiwara H et al. Cloning and properties of a novel natriuretic peptide receptor, NPR-D. European Journal of Biochemistry. 1995;233(1):102-109.
Kashiwagi, M. ; Katafuchi, T. ; Kato, A. ; Inuyama, H. ; Ito, T. ; Hagiwara, H. ; Takei, Y. ; Hirose, S. / Cloning and properties of a novel natriuretic peptide receptor, NPR-D. In: European Journal of Biochemistry. 1995 ; Vol. 233, No. 1. pp. 102-109.
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