Attenuated desensitization of β-adrenergic receptor by water-soluble N-nitrosamines that induce S-nitrosylation without NO release

Noriko Makita, Yoji Kabasawa, Yuko Otani, Firman, Junichiro Sato, Makiko Hashimoto, Michio Nakaya, Hiroaki Nishihara, Masaomi Nangaku, Hitoshi Kurose, Tomohiko Ohwada, Taroh Iiri

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Rationale: The clinical problem of loss of β-adrenergic receptor (β-AR) response, both in the pathogenesis of heart failure and during therapeutic application of β-agonists, is attributable, at least in part, to desensitization, internalization, and downregulation of the receptors. In the regulation of β-AR signaling, G protein-coupled receptor kinase 2 (GRK2) primarily phosphorylates agonist-occupied β-ARs, and this modification promotes desensitization, internalization, and downregulation of β-ARs. It has been demonstrated that GRK2 is inhibited by its S-nitrosylation. However, compounds that induce S-nitrosylation, such as S-nitrosoglutathione, simultaneously generate NO, which has been demonstrated to operate for cardiovascular protection. Objective: We examine whether S-nitrosylation without NO generation inhibits desensitization of β2-AR by GRK2. We thus aim to synthesize compounds that specifically induce S-nitrosylation. Methods and Results: We have developed water-soluble N-nitrosamines that have S-nitrosylating activity but lack NO-generating activity. These compounds, at least partly, rescue β-AR from desensitization in HEK 293 cells expressing FLAG-tagged human β2-AR and in rat cardiac myocytes. They inhibit isoproterenol-dependent phosphorylation and internalization of β2-AR. Indeed, they nitrosylate GRK2 in vitro and in cells, and their S-nitrosylation of GRK2 likely underlies their inhibition of β2-AR desensitization. Conclusions: Compounds that induce S-nitrosylation without NO release inhibit GRK2 and attenuate β2-AR desensitization. Developing water-soluble drugs that specifically induce S-nitrosylation may be a promising therapeutic strategy for heart failure.

Original languageEnglish
Pages (from-to)327-334
Number of pages8
JournalCirculation research
Volume112
Issue number2
DOIs
Publication statusPublished - Jan 18 2013

Fingerprint

G-Protein-Coupled Receptor Kinase 2
Nitrosamines
Adrenergic Receptors
Water
Down-Regulation
Heart Failure
S-Nitrosoglutathione
HEK293 Cells
Isoproterenol
Cardiac Myocytes
Phosphorylation
Therapeutics
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Attenuated desensitization of β-adrenergic receptor by water-soluble N-nitrosamines that induce S-nitrosylation without NO release. / Makita, Noriko; Kabasawa, Yoji; Otani, Yuko; Firman; Sato, Junichiro; Hashimoto, Makiko; Nakaya, Michio; Nishihara, Hiroaki; Nangaku, Masaomi; Kurose, Hitoshi; Ohwada, Tomohiko; Iiri, Taroh.

In: Circulation research, Vol. 112, No. 2, 18.01.2013, p. 327-334.

Research output: Contribution to journalArticle

Makita, N, Kabasawa, Y, Otani, Y, Firman, Sato, J, Hashimoto, M, Nakaya, M, Nishihara, H, Nangaku, M, Kurose, H, Ohwada, T & Iiri, T 2013, 'Attenuated desensitization of β-adrenergic receptor by water-soluble N-nitrosamines that induce S-nitrosylation without NO release', Circulation research, vol. 112, no. 2, pp. 327-334. https://doi.org/10.1161/CIRCRESAHA.112.277665
Makita, Noriko ; Kabasawa, Yoji ; Otani, Yuko ; Firman ; Sato, Junichiro ; Hashimoto, Makiko ; Nakaya, Michio ; Nishihara, Hiroaki ; Nangaku, Masaomi ; Kurose, Hitoshi ; Ohwada, Tomohiko ; Iiri, Taroh. / Attenuated desensitization of β-adrenergic receptor by water-soluble N-nitrosamines that induce S-nitrosylation without NO release. In: Circulation research. 2013 ; Vol. 112, No. 2. pp. 327-334.
@article{b96497f8d40e431695b0d76922cc6013,
title = "Attenuated desensitization of β-adrenergic receptor by water-soluble N-nitrosamines that induce S-nitrosylation without NO release",
abstract = "Rationale: The clinical problem of loss of β-adrenergic receptor (β-AR) response, both in the pathogenesis of heart failure and during therapeutic application of β-agonists, is attributable, at least in part, to desensitization, internalization, and downregulation of the receptors. In the regulation of β-AR signaling, G protein-coupled receptor kinase 2 (GRK2) primarily phosphorylates agonist-occupied β-ARs, and this modification promotes desensitization, internalization, and downregulation of β-ARs. It has been demonstrated that GRK2 is inhibited by its S-nitrosylation. However, compounds that induce S-nitrosylation, such as S-nitrosoglutathione, simultaneously generate NO, which has been demonstrated to operate for cardiovascular protection. Objective: We examine whether S-nitrosylation without NO generation inhibits desensitization of β2-AR by GRK2. We thus aim to synthesize compounds that specifically induce S-nitrosylation. Methods and Results: We have developed water-soluble N-nitrosamines that have S-nitrosylating activity but lack NO-generating activity. These compounds, at least partly, rescue β-AR from desensitization in HEK 293 cells expressing FLAG-tagged human β2-AR and in rat cardiac myocytes. They inhibit isoproterenol-dependent phosphorylation and internalization of β2-AR. Indeed, they nitrosylate GRK2 in vitro and in cells, and their S-nitrosylation of GRK2 likely underlies their inhibition of β2-AR desensitization. Conclusions: Compounds that induce S-nitrosylation without NO release inhibit GRK2 and attenuate β2-AR desensitization. Developing water-soluble drugs that specifically induce S-nitrosylation may be a promising therapeutic strategy for heart failure.",
author = "Noriko Makita and Yoji Kabasawa and Yuko Otani and Firman and Junichiro Sato and Makiko Hashimoto and Michio Nakaya and Hiroaki Nishihara and Masaomi Nangaku and Hitoshi Kurose and Tomohiko Ohwada and Taroh Iiri",
year = "2013",
month = "1",
day = "18",
doi = "10.1161/CIRCRESAHA.112.277665",
language = "English",
volume = "112",
pages = "327--334",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "Lippincott Williams and Wilkins",
number = "2",

}

TY - JOUR

T1 - Attenuated desensitization of β-adrenergic receptor by water-soluble N-nitrosamines that induce S-nitrosylation without NO release

AU - Makita, Noriko

AU - Kabasawa, Yoji

AU - Otani, Yuko

AU - Firman,

AU - Sato, Junichiro

AU - Hashimoto, Makiko

AU - Nakaya, Michio

AU - Nishihara, Hiroaki

AU - Nangaku, Masaomi

AU - Kurose, Hitoshi

AU - Ohwada, Tomohiko

AU - Iiri, Taroh

PY - 2013/1/18

Y1 - 2013/1/18

N2 - Rationale: The clinical problem of loss of β-adrenergic receptor (β-AR) response, both in the pathogenesis of heart failure and during therapeutic application of β-agonists, is attributable, at least in part, to desensitization, internalization, and downregulation of the receptors. In the regulation of β-AR signaling, G protein-coupled receptor kinase 2 (GRK2) primarily phosphorylates agonist-occupied β-ARs, and this modification promotes desensitization, internalization, and downregulation of β-ARs. It has been demonstrated that GRK2 is inhibited by its S-nitrosylation. However, compounds that induce S-nitrosylation, such as S-nitrosoglutathione, simultaneously generate NO, which has been demonstrated to operate for cardiovascular protection. Objective: We examine whether S-nitrosylation without NO generation inhibits desensitization of β2-AR by GRK2. We thus aim to synthesize compounds that specifically induce S-nitrosylation. Methods and Results: We have developed water-soluble N-nitrosamines that have S-nitrosylating activity but lack NO-generating activity. These compounds, at least partly, rescue β-AR from desensitization in HEK 293 cells expressing FLAG-tagged human β2-AR and in rat cardiac myocytes. They inhibit isoproterenol-dependent phosphorylation and internalization of β2-AR. Indeed, they nitrosylate GRK2 in vitro and in cells, and their S-nitrosylation of GRK2 likely underlies their inhibition of β2-AR desensitization. Conclusions: Compounds that induce S-nitrosylation without NO release inhibit GRK2 and attenuate β2-AR desensitization. Developing water-soluble drugs that specifically induce S-nitrosylation may be a promising therapeutic strategy for heart failure.

AB - Rationale: The clinical problem of loss of β-adrenergic receptor (β-AR) response, both in the pathogenesis of heart failure and during therapeutic application of β-agonists, is attributable, at least in part, to desensitization, internalization, and downregulation of the receptors. In the regulation of β-AR signaling, G protein-coupled receptor kinase 2 (GRK2) primarily phosphorylates agonist-occupied β-ARs, and this modification promotes desensitization, internalization, and downregulation of β-ARs. It has been demonstrated that GRK2 is inhibited by its S-nitrosylation. However, compounds that induce S-nitrosylation, such as S-nitrosoglutathione, simultaneously generate NO, which has been demonstrated to operate for cardiovascular protection. Objective: We examine whether S-nitrosylation without NO generation inhibits desensitization of β2-AR by GRK2. We thus aim to synthesize compounds that specifically induce S-nitrosylation. Methods and Results: We have developed water-soluble N-nitrosamines that have S-nitrosylating activity but lack NO-generating activity. These compounds, at least partly, rescue β-AR from desensitization in HEK 293 cells expressing FLAG-tagged human β2-AR and in rat cardiac myocytes. They inhibit isoproterenol-dependent phosphorylation and internalization of β2-AR. Indeed, they nitrosylate GRK2 in vitro and in cells, and their S-nitrosylation of GRK2 likely underlies their inhibition of β2-AR desensitization. Conclusions: Compounds that induce S-nitrosylation without NO release inhibit GRK2 and attenuate β2-AR desensitization. Developing water-soluble drugs that specifically induce S-nitrosylation may be a promising therapeutic strategy for heart failure.

UR - http://www.scopus.com/inward/record.url?scp=84872876881&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84872876881&partnerID=8YFLogxK

U2 - 10.1161/CIRCRESAHA.112.277665

DO - 10.1161/CIRCRESAHA.112.277665

M3 - Article

C2 - 23212582

AN - SCOPUS:84872876881

VL - 112

SP - 327

EP - 334

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 2

ER -