Uncovering kappa-opioid receptor agonist-induced PAK1/2 phosphorylation by quantitative phosphoproteomics

Jianhong Wu; Qinghui Jiang; Hongwen Zhu; Yanting Zhou; Dayun Lu; Xing Liu; Xiangling Chen; Jie Chen; Yujun Wang; Jinggen Liu; Rentao Song; Ruimin Huang; Hu Zhou

doi:10.1016/j.bbrc.2019.06.111

Uncovering kappa-opioid receptor agonist-induced PAK1/2 phosphorylation by quantitative phosphoproteomics

Biochem Biophys Res Commun. 2019 Aug 13;516(1):320-326. doi: 10.1016/j.bbrc.2019.06.111. Epub 2019 Jun 28.

Authors

Jianhong Wu¹, Qinghui Jiang², Hongwen Zhu³, Yanting Zhou³, Dayun Lu², Xing Liu³, Xiangling Chen², Jie Chen³, Yujun Wang², Jinggen Liu², Rentao Song⁴, Ruimin Huang⁵, Hu Zhou⁶

Affiliations

¹ School of Life Sciences, Shanghai University, Shanghai, China; Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
² Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China; University of Chinese Academy of Sciences, Beijing, China.
³ Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
⁴ School of Life Sciences, Shanghai University, Shanghai, China.
⁵ Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China; University of Chinese Academy of Sciences, Beijing, China. Electronic address: rmhuang@simm.ac.cn.
⁶ Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China; University of Chinese Academy of Sciences, Beijing, China; E-institute of Shanghai Municipal Education Committee, Shanghai University of Traditional Chinese Medicine, Shanghai, China. Electronic address: zhouhu@simm.ac.cn.

PMID: 31256935
DOI: 10.1016/j.bbrc.2019.06.111

Abstract

Kappa-opioid receptor (KOR) is a member of G-protein coupled receptors (GPCRs) expressed in serotonergic neurons and neuronal terminals. The involvement of KOR ligands in nociception, diuresis, emotion, cognition, and immune system has been extensively studied. Omics-based methods are preferable to understand the signaling cascade after KOR activation in a systematic manner. In this study, an in-depth quantitative phosphoproteomic analysis resulted in 305 phosphosites, which were significantly changed in three KOR-overexpressed cells upon treatment with two KOR agonists. The subsequent substrate-kinase prediction analysis revealed that 18 potential kinases might be activated under stimulation of the agonists. We found that phosphorylation of PAK1/2 (p21-activated kinase 1/2) was induced by KOR agonists, resulting in reduced actin stress fibers and cytoskeletal reorganization. In summary, this quantitative phosphoproteomics-based research studied the downstream phosphorylation events upon KOR activation, which may shed light on the investigations of KOR signaling pathway and targeted therapy for KOR-related diseases.

Keywords: Dimethyl labeling; Kappa-opioid receptor; Kinase-substrate network; Quantitative phosphoproteomics; p21-activated kinases.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Enzyme Activation / drug effects
Enzyme Activators / pharmacology*
HEK293 Cells
Humans
Phosphorylation / drug effects*
Proteomics
Receptors, Opioid, kappa / agonists*
Receptors, Opioid, kappa / metabolism
p21-Activated Kinases / metabolism*

Substances

Enzyme Activators
Receptors, Opioid, kappa
PAK1 protein, human
PAK2 protein, human
p21-Activated Kinases