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Characterization of a rat type 2 angiotensin II receptor stably expressed in 293 cells.

Publication ,  Journal Article
Mukoyama, M; Horiuchi, M; Nakajima, M; Pratt, RE; Dzau, VJ
Published in: Mol Cell Endocrinol
July 1995

A cDNA clone for the rat type 2 (AT2) angiotensin II receptor was stably transfected into human embryonic kidney 293 cells. Binding characteristics of CGP42112A (Kd = 0.18 nM, Bmax = 10.8 pmol/mg protein) and ligand specificity were indistinguishable from those obtained with the whole rat fetus and with transiently transfected COS-7 cells. Non-hydrolyzable guanine nucleotide analogs did not affect the ligand binding curve; interestingly, the guanine nucleotide analog's effect was observed in the presence of sulfhydryl reducing agent, suggesting that a certain redox condition may affect G protein coupling to this receptor. Using the established cell line, several second messenger systems were assessed. None of cAMP levels, cGMP levels, arachidonic acid release, or phosphotyrosine phosphatase activity was affected by angiotensin II stimulation of this receptor. Furthermore, the AT2 receptor did not undergo agonist-stimulated internalization. These results using the cloned receptor suggest that the transfected AT2 receptor fails to effectively couple to the major G protein-mediated signaling mechanisms and ligand-activated internalization in transfected 293 cells.

Duke Scholars

Published In

Mol Cell Endocrinol

DOI

ISSN

0303-7207

Publication Date

July 1995

Volume

112

Issue

1

Start / End Page

61 / 68

Location

Ireland

Related Subject Headings

  • Transfection
  • Recombinant Proteins
  • Receptors, Angiotensin
  • Rats
  • Oxidation-Reduction
  • Oligopeptides
  • Kidney
  • Iodine Radioisotopes
  • Humans
  • Guanine Nucleotides
 

Citation

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Mukoyama, M., Horiuchi, M., Nakajima, M., Pratt, R. E., & Dzau, V. J. (1995). Characterization of a rat type 2 angiotensin II receptor stably expressed in 293 cells. Mol Cell Endocrinol, 112(1), 61–68. https://doi.org/10.1016/0303-7207(95)03586-v
Mukoyama, M., M. Horiuchi, M. Nakajima, R. E. Pratt, and V. J. Dzau. “Characterization of a rat type 2 angiotensin II receptor stably expressed in 293 cells.Mol Cell Endocrinol 112, no. 1 (July 1995): 61–68. https://doi.org/10.1016/0303-7207(95)03586-v.
Mukoyama M, Horiuchi M, Nakajima M, Pratt RE, Dzau VJ. Characterization of a rat type 2 angiotensin II receptor stably expressed in 293 cells. Mol Cell Endocrinol. 1995 Jul;112(1):61–8.
Mukoyama, M., et al. “Characterization of a rat type 2 angiotensin II receptor stably expressed in 293 cells.Mol Cell Endocrinol, vol. 112, no. 1, July 1995, pp. 61–68. Pubmed, doi:10.1016/0303-7207(95)03586-v.
Mukoyama M, Horiuchi M, Nakajima M, Pratt RE, Dzau VJ. Characterization of a rat type 2 angiotensin II receptor stably expressed in 293 cells. Mol Cell Endocrinol. 1995 Jul;112(1):61–68.
Journal cover image

Published In

Mol Cell Endocrinol

DOI

ISSN

0303-7207

Publication Date

July 1995

Volume

112

Issue

1

Start / End Page

61 / 68

Location

Ireland

Related Subject Headings

  • Transfection
  • Recombinant Proteins
  • Receptors, Angiotensin
  • Rats
  • Oxidation-Reduction
  • Oligopeptides
  • Kidney
  • Iodine Radioisotopes
  • Humans
  • Guanine Nucleotides