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Role of basic amino acids of the human angiotensin type 1 receptor in the binding of the non-peptide antagonist candesartan

Department of Molecular and Biochemical Pharmacology, Institute for Molecular Biology and Biotechnology, Free University of Brussels (VUB), B-1640 Sint-Genesius Rode, Belgium, gvauquel@ vub.ac.be

Frederik LP Fierens

Department of Molecular and Biochemical Pharmacology, Institute for Molecular Biology and Biotechnology, Free University of Brussels (VUB), B-1640 Sint-Genesius Rode, Belgium

Zsuzsanna Gáborik

Department of Physiology, Semmelweis University Medical School, H-1444 Budapest, PO Box 259, Hungary

Tam Le Minh

Department of Molecular and Biochemical Pharmacology, Institute for Molecular Biology and Biotechnology, Free University of Brussels (VUB), B-1640 Sint-Genesius Rode, Belgium

Jean-Paul De Backer

Department of Molecular and Biochemical Pharmacology, Institute for Molecular Biology and Biotechnology, Free University of Brussels (VUB), B-1640 Sint-Genesius Rode, Belgium

László Hunyady

Department of Physiology, Semmelweis University Medical School, H-1444 Budapest, PO Box 259, Hungary

Patrick ML Vanderheyden

Department of Molecular and Biochemical Pharmacology, Institute for Molecular Biology and Biotechnology, Free University of Brussels (VUB), B-1640 Sint-Genesius Rode, Belgium

To explain the insurmountable/long-lasting binding of biphenyltetrazole-containing AT₁-receptor antagonists such as candesartan, to the human angiotensin II type 1-receptor, a model is proposed in which the basic amino acids Lys¹⁹⁹ and Arg ¹⁶⁷ of the receptor interact respectively with the carboxylate and the tetrazole group of the antagonists. To validate this model, we have investigated the impact of substitution of Lys¹⁹⁹ by Ala or Gln and of Arg¹⁶⁷ by Ala on the binding properties of [³H]candesartan and on competition binding by candesartan, EXP3174, irbesartan, losartan, angiotensin II (Ang II) and [Sar¹-Ile⁸]angiotensin. Our results indicate that both amino acids play an important role in the AT₁-receptor ligand binding. Whereas the negative charge of Lys ¹⁹⁹ is involved in an ionic bond with the end-standing carboxylate group of the peptide ligands, its polarity also contributes to the non-peptide antagonist binding. Substitution of Arg¹⁶⁷ by Ala completely abolished [³H]Ang II, as well as [³H] candesartan, binding. Whereas these results are in line with the proposed model, it cannot be excluded that both amino acid residues are important for the structural integrity of the AT₁-receptor with respect to its ligand binding properties.

Key Words: CHO cells • angiotensin II • human AT1-receptor • mutation • candesartan • insurmountable • non-peptide antagonist

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Journal of Renin-Angiotensin-Aldosterone System, Vol. 2, No. 1 suppl, S32-S36 (2001)
DOI: 10.1177/14703203010020010501


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This Article Abstract Free Full Text (Free PDF) Free Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Vauquelin, G. Articles by Vanderheyden, P. M. Search for Related Content PubMed Articles by Vauquelin, G. Articles by Vanderheyden, P. M. Social Bookmarking                         What's this?