Two C-terminal amino acids, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of opioid receptor-like 1 receptors

Hung Li Wang, Yo Li Kuo, Chia Yu Hsu, Pei Chen Huang, Allen H. Li, An Hsun Chou, Tu Hsueh Yeh, Ying Ling Chen

研究成果: 雜誌貢獻文章

5 引文 (Scopus)

摘要

Various cellular signaling pathways induced by nociceptin activation of ORL1 (opioid receptor-like 1 receptor) develop homologous desensitization. Multiple lines of evidence suggest that agonist-induced phosphorylation of serine (Ser)/threonine (Thr) residues at intracellular carboxyl tail leads to homologous desensitization of G protein-coupled receptors. In the present study, we investigated the functional role played by C-terminal Ser/Thr residues in agonist-induced desensitization and phosphorylation of ORL1. In HEK 293 cells expressing wild-type ORL1 and ORL1(CΔ21), which lacks C-terminal 21 amino acids, nociceptin inhibition of adenylate cyclase activity exhibited homologous desensitization after 1 h pretreatment of nociceptin. In contrast, ORL1(CΔ34), which differs with ORL1(CΔ21) by lacking C-terminal Ser334, Ser335 and Ser343 residues, failed to develop agonist-induced desensitization. Point mutant (S343A) ORL1 underwent homologous desensitization after nociceptin pretreatment. Substituting Ser 334 or Ser335 with alanine greatly impaired nociceptin-induced ORL1 desensitization. In HEK 293 cells expressing double mutant (S334A/S335A) ORL1, nociceptin pretreatment failed to significantly affect the efficacy and potency by which nociceptin inhibits forskolin-stimulated cAMP formation. Mutation of Ser334 and Ser 335 also greatly reduced nociceptin-induced ORL1 phosphorylation. These results suggest that two C-terminal serine residues, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of ORL1.
原文英語
頁(從 - 到)670-678
頁數9
期刊Cellular Signalling
18
發行號5
DOIs
出版狀態已發佈 - 五月 1 2006

指紋

Opioid Receptors
Phosphorylation
Amino Acids
Serine
HEK293 Cells
Threonine
Colforsin
G-Protein-Coupled Receptors
nociceptin
Adenylyl Cyclases
Alanine
Mutation

ASJC Scopus subject areas

  • Cell Biology

引用此文

Two C-terminal amino acids, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of opioid receptor-like 1 receptors. / Wang, Hung Li; Kuo, Yo Li; Hsu, Chia Yu; Huang, Pei Chen; Li, Allen H.; Chou, An Hsun; Yeh, Tu Hsueh; Chen, Ying Ling.

於: Cellular Signalling, 卷 18, 編號 5, 01.05.2006, p. 670-678.

研究成果: 雜誌貢獻文章

Wang, Hung Li ; Kuo, Yo Li ; Hsu, Chia Yu ; Huang, Pei Chen ; Li, Allen H. ; Chou, An Hsun ; Yeh, Tu Hsueh ; Chen, Ying Ling. / Two C-terminal amino acids, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of opioid receptor-like 1 receptors. 於: Cellular Signalling. 2006 ; 卷 18, 編號 5. 頁 670-678.
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title = "Two C-terminal amino acids, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of opioid receptor-like 1 receptors",
abstract = "Various cellular signaling pathways induced by nociceptin activation of ORL1 (opioid receptor-like 1 receptor) develop homologous desensitization. Multiple lines of evidence suggest that agonist-induced phosphorylation of serine (Ser)/threonine (Thr) residues at intracellular carboxyl tail leads to homologous desensitization of G protein-coupled receptors. In the present study, we investigated the functional role played by C-terminal Ser/Thr residues in agonist-induced desensitization and phosphorylation of ORL1. In HEK 293 cells expressing wild-type ORL1 and ORL1(CΔ21), which lacks C-terminal 21 amino acids, nociceptin inhibition of adenylate cyclase activity exhibited homologous desensitization after 1 h pretreatment of nociceptin. In contrast, ORL1(CΔ34), which differs with ORL1(CΔ21) by lacking C-terminal Ser334, Ser335 and Ser343 residues, failed to develop agonist-induced desensitization. Point mutant (S343A) ORL1 underwent homologous desensitization after nociceptin pretreatment. Substituting Ser 334 or Ser335 with alanine greatly impaired nociceptin-induced ORL1 desensitization. In HEK 293 cells expressing double mutant (S334A/S335A) ORL1, nociceptin pretreatment failed to significantly affect the efficacy and potency by which nociceptin inhibits forskolin-stimulated cAMP formation. Mutation of Ser334 and Ser 335 also greatly reduced nociceptin-induced ORL1 phosphorylation. These results suggest that two C-terminal serine residues, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of ORL1.",
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author = "Wang, {Hung Li} and Kuo, {Yo Li} and Hsu, {Chia Yu} and Huang, {Pei Chen} and Li, {Allen H.} and Chou, {An Hsun} and Yeh, {Tu Hsueh} and Chen, {Ying Ling}",
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T1 - Two C-terminal amino acids, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of opioid receptor-like 1 receptors

AU - Wang, Hung Li

AU - Kuo, Yo Li

AU - Hsu, Chia Yu

AU - Huang, Pei Chen

AU - Li, Allen H.

AU - Chou, An Hsun

AU - Yeh, Tu Hsueh

AU - Chen, Ying Ling

PY - 2006/5/1

Y1 - 2006/5/1

N2 - Various cellular signaling pathways induced by nociceptin activation of ORL1 (opioid receptor-like 1 receptor) develop homologous desensitization. Multiple lines of evidence suggest that agonist-induced phosphorylation of serine (Ser)/threonine (Thr) residues at intracellular carboxyl tail leads to homologous desensitization of G protein-coupled receptors. In the present study, we investigated the functional role played by C-terminal Ser/Thr residues in agonist-induced desensitization and phosphorylation of ORL1. In HEK 293 cells expressing wild-type ORL1 and ORL1(CΔ21), which lacks C-terminal 21 amino acids, nociceptin inhibition of adenylate cyclase activity exhibited homologous desensitization after 1 h pretreatment of nociceptin. In contrast, ORL1(CΔ34), which differs with ORL1(CΔ21) by lacking C-terminal Ser334, Ser335 and Ser343 residues, failed to develop agonist-induced desensitization. Point mutant (S343A) ORL1 underwent homologous desensitization after nociceptin pretreatment. Substituting Ser 334 or Ser335 with alanine greatly impaired nociceptin-induced ORL1 desensitization. In HEK 293 cells expressing double mutant (S334A/S335A) ORL1, nociceptin pretreatment failed to significantly affect the efficacy and potency by which nociceptin inhibits forskolin-stimulated cAMP formation. Mutation of Ser334 and Ser 335 also greatly reduced nociceptin-induced ORL1 phosphorylation. These results suggest that two C-terminal serine residues, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of ORL1.

AB - Various cellular signaling pathways induced by nociceptin activation of ORL1 (opioid receptor-like 1 receptor) develop homologous desensitization. Multiple lines of evidence suggest that agonist-induced phosphorylation of serine (Ser)/threonine (Thr) residues at intracellular carboxyl tail leads to homologous desensitization of G protein-coupled receptors. In the present study, we investigated the functional role played by C-terminal Ser/Thr residues in agonist-induced desensitization and phosphorylation of ORL1. In HEK 293 cells expressing wild-type ORL1 and ORL1(CΔ21), which lacks C-terminal 21 amino acids, nociceptin inhibition of adenylate cyclase activity exhibited homologous desensitization after 1 h pretreatment of nociceptin. In contrast, ORL1(CΔ34), which differs with ORL1(CΔ21) by lacking C-terminal Ser334, Ser335 and Ser343 residues, failed to develop agonist-induced desensitization. Point mutant (S343A) ORL1 underwent homologous desensitization after nociceptin pretreatment. Substituting Ser 334 or Ser335 with alanine greatly impaired nociceptin-induced ORL1 desensitization. In HEK 293 cells expressing double mutant (S334A/S335A) ORL1, nociceptin pretreatment failed to significantly affect the efficacy and potency by which nociceptin inhibits forskolin-stimulated cAMP formation. Mutation of Ser334 and Ser 335 also greatly reduced nociceptin-induced ORL1 phosphorylation. These results suggest that two C-terminal serine residues, Ser334 and Ser335, are required for homologous desensitization and agonist-induced phosphorylation of ORL1.

KW - Adenylate cyclase

KW - Agonist-induced phosphorylation

KW - HEK 293 cells

KW - Homologous desensitization

KW - Nociceptin

KW - Opioid receptor-like 1 receptor

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U2 - 10.1016/j.cellsig.2005.06.009

DO - 10.1016/j.cellsig.2005.06.009

M3 - Article

C2 - 16019191

AN - SCOPUS:30944460115

VL - 18

SP - 670

EP - 678

JO - Cellular Signalling

JF - Cellular Signalling

SN - 0898-6568

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