β1-selective agonist (-)-1-(3,4-dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanol [(-)-RO363] differentially interacts with key amino acids responsible for β1-selective binding in resting and active states

Yoshiyuki Sugimoto, Reiko Fujisawa, Ryuji Tanimura, Anne Laure Lattion, Susanna Cotecchia, Gozoh Tsujimoto, Taku Nagao, Hitoshi Kurose

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

(-)-1-(3,4-Dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2- propanol [(-)-RO363] is a highly selective β1-adrenergic receptor (β1AR) agonist. To study the binding site of β1-selective agonist, chimeric β12 and Ala-substituted β1ARs were constructed. Several key residues of β1AR [Leu110 and Thr117 in transmembrane domain (TMD) 2], and Phe359 in TMD 7] were found to be responsible for β1-selective binding of (-)-RO363, as determined by competitive binding. Based on these results, we built a three-dimensional model of the binding domain for (-)-RO363. The model indicated that TMD 2 and TMD 7 of β1AR form a binding pocket; the methoxyphenyl group of N-substituent of (-)-RO363 seems to locate within the cavity surrounded by Leu110, Thr117, and Phe359. The amino acids Leu110 and Phe359 interact with the phenyl ring of (-)-RO363, whereas Thr117 forms hydrogen bond with the methoxy group of (-)-RO363. To examine the interaction of these residues with β1 AR in an active state, each of the amino acids was changed to Ala in a constitutively active (CA)-β1AR mutant. The degree of decrease in the affinity of CA-β1AR for (-)-RO363 was essentially the same as that of wild-type β1AR when mutated at Leu110 and Thr117. However, the affinity was decreased in Ala-substituted mutant of Phe359 compared with that of wild-type β1AR. These results indicated that Leu110 and Thr117 are necessary for the initial binding of (-)-RO363 with β1-selectivity, and interaction of Phe359 with the N-substituent of (-)-RO363 in an active state is stronger than in the resting state.

Original languageEnglish
Pages (from-to)51-58
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume301
Issue number1
DOIs
Publication statusPublished - Apr 6 2002
Externally publishedYes

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2-Propanol
Amino Acids
Adrenergic Receptors
Ro 363
1,21-diamino-4,9,13,18-tetraazahenicosane
Adrenergic Agonists
Competitive Binding
Hydrogen
Binding Sites

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

β1-selective agonist (-)-1-(3,4-dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanol [(-)-RO363] differentially interacts with key amino acids responsible for β1-selective binding in resting and active states. / Sugimoto, Yoshiyuki; Fujisawa, Reiko; Tanimura, Ryuji; Lattion, Anne Laure; Cotecchia, Susanna; Tsujimoto, Gozoh; Nagao, Taku; Kurose, Hitoshi.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 301, No. 1, 06.04.2002, p. 51-58.

Research output: Contribution to journalArticle

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title = "β1-selective agonist (-)-1-(3,4-dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanol [(-)-RO363] differentially interacts with key amino acids responsible for β1-selective binding in resting and active states",
abstract = "(-)-1-(3,4-Dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2- propanol [(-)-RO363] is a highly selective β1-adrenergic receptor (β1AR) agonist. To study the binding site of β1-selective agonist, chimeric β1/β2 and Ala-substituted β1ARs were constructed. Several key residues of β1AR [Leu110 and Thr117 in transmembrane domain (TMD) 2], and Phe359 in TMD 7] were found to be responsible for β1-selective binding of (-)-RO363, as determined by competitive binding. Based on these results, we built a three-dimensional model of the binding domain for (-)-RO363. The model indicated that TMD 2 and TMD 7 of β1AR form a binding pocket; the methoxyphenyl group of N-substituent of (-)-RO363 seems to locate within the cavity surrounded by Leu110, Thr117, and Phe359. The amino acids Leu110 and Phe359 interact with the phenyl ring of (-)-RO363, whereas Thr117 forms hydrogen bond with the methoxy group of (-)-RO363. To examine the interaction of these residues with β1 AR in an active state, each of the amino acids was changed to Ala in a constitutively active (CA)-β1AR mutant. The degree of decrease in the affinity of CA-β1AR for (-)-RO363 was essentially the same as that of wild-type β1AR when mutated at Leu110 and Thr117. However, the affinity was decreased in Ala-substituted mutant of Phe359 compared with that of wild-type β1AR. These results indicated that Leu110 and Thr117 are necessary for the initial binding of (-)-RO363 with β1-selectivity, and interaction of Phe359 with the N-substituent of (-)-RO363 in an active state is stronger than in the resting state.",
author = "Yoshiyuki Sugimoto and Reiko Fujisawa and Ryuji Tanimura and Lattion, {Anne Laure} and Susanna Cotecchia and Gozoh Tsujimoto and Taku Nagao and Hitoshi Kurose",
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T1 - β1-selective agonist (-)-1-(3,4-dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanol [(-)-RO363] differentially interacts with key amino acids responsible for β1-selective binding in resting and active states

AU - Sugimoto, Yoshiyuki

AU - Fujisawa, Reiko

AU - Tanimura, Ryuji

AU - Lattion, Anne Laure

AU - Cotecchia, Susanna

AU - Tsujimoto, Gozoh

AU - Nagao, Taku

AU - Kurose, Hitoshi

PY - 2002/4/6

Y1 - 2002/4/6

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AB - (-)-1-(3,4-Dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2- propanol [(-)-RO363] is a highly selective β1-adrenergic receptor (β1AR) agonist. To study the binding site of β1-selective agonist, chimeric β1/β2 and Ala-substituted β1ARs were constructed. Several key residues of β1AR [Leu110 and Thr117 in transmembrane domain (TMD) 2], and Phe359 in TMD 7] were found to be responsible for β1-selective binding of (-)-RO363, as determined by competitive binding. Based on these results, we built a three-dimensional model of the binding domain for (-)-RO363. The model indicated that TMD 2 and TMD 7 of β1AR form a binding pocket; the methoxyphenyl group of N-substituent of (-)-RO363 seems to locate within the cavity surrounded by Leu110, Thr117, and Phe359. The amino acids Leu110 and Phe359 interact with the phenyl ring of (-)-RO363, whereas Thr117 forms hydrogen bond with the methoxy group of (-)-RO363. To examine the interaction of these residues with β1 AR in an active state, each of the amino acids was changed to Ala in a constitutively active (CA)-β1AR mutant. The degree of decrease in the affinity of CA-β1AR for (-)-RO363 was essentially the same as that of wild-type β1AR when mutated at Leu110 and Thr117. However, the affinity was decreased in Ala-substituted mutant of Phe359 compared with that of wild-type β1AR. These results indicated that Leu110 and Thr117 are necessary for the initial binding of (-)-RO363 with β1-selectivity, and interaction of Phe359 with the N-substituent of (-)-RO363 in an active state is stronger than in the resting state.

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