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The β1-adrenergic receptor mediates extracellular signal-regulated kinase activation via Gαs

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Abstract

β-Adrenergic receptors can activate extracellular signal-regulated kinases (ERKs) via different mechanisms. In this study, we investigated the molecular mechanism of β1-adrenergic receptor (β1AR)-mediated ERK activation in African green monkey kidney COS-7 cells. Treatment of cells with isoproterenol (ISO), a β1AR selective agonist, induced phosphorylation of ERK1/2 in a dose-dependent manner. ISO-stimulated ERK phosphorylation was not influenced by the Gβγ inhibitor, βAR kinase carboxyl terminal (βARKct) or by the Gi inhibitor, pertussis toxin (PTX), but it was clearly abolished via inhibition of protein kinase A (PKA) with H89, or of mitogen-activated protein kinase kinase (MEK1) with PD98059, revealing that the Gαs subunit is involved in ERK regulation through the PKA/MEK1 pathway. We also tested the effect of the adenylate cyclase activator forskolin on ERK activation, and the result was identical to that of ISO stimulation. Moreover, pretreatment with the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor AG1478 or with the Src tyrosine kinase inhibitor PP2 did not affect ERK activation. These observations suggest a mechanism of β1AR-mediated ERK activity that involves the Gαs subunit, but not EGFR or Src tyrosine kinase.

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Abbreviations

COS-7:

African green monkey kidney cells

DMEM:

Dulbecco’s modified Eagle’s medium

ISO:

Isoproterenol

β1AR:

β1-Adrenergic receptor

GPCR:

G protein-coupled receptor

G protein:

GTP binding regulatory protein

MAP:

Mitogen-activated protein

ERK:

The mitogen-activated protein kinases extracellular signal-regulated kinase

βARK:

β-Adrenergic receptor kinase

CT:

Carboxyl terminal

FSK:

Forskolin

AC:

Adenylate cyclase

PTX:

Pertussis toxin

cAMP:

Cyclic AMP

PKA:

Protein kinase A

PKC:

Protein kinase C

H89:

N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide

PP2:

4-Amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo [3,4-d]pyrimidine

MEK1:

Mitogen-activated ERK kinase 1

PD98059:

2-(2-Amino-3-methoxyphenol)-oxanaphthalen-4-one

EGFR:

Epidermal growth factor receptor

AG1478:

2-(2-Amino-3-methoxyphenyl)-oxanaphthalen-4-one, and 4-(3-chloroanilino)-6,7-dimethoxyquinazoline

SDS:

Sodium dodecyl sulfate

PAGE:

Polyacrylamide gel electrophoresis

PSD-95:

Postsynaptic density-95

MAGI-2:

Membrane-associated guanylate kinase inverted-2

CNrasGEF:

cAMP-dependent guanine nucleotide exchange factor

GIPC:

GAIP-interacting protein carboxyl terminus

CAL:

Cystic fibrosis transmembrane conductance regulator-associated ligand

MAGUK:

Membrane-associated guanylate kinase

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Acknowledgments

This work was supported by National Natural Science Foundation of the People’s Republic of China (No: 30572183 and 30772573), the New Century Excellent Talents in University of China (NCET-06-0184), Research Fund for the Doctoral Program of Higher Education of China (20060025011), Foundation of Beijing Educational Committee (KZ200610025013), Natural Science Foundation of Beijing (No: 7082009) and Scientific Research Foundation for Returned Scholars, Ministry of Education of China awarded to J. He, and fund for Excellent Talents in Beijing (20071D0501800253) awarded to JF Zheng. The manuscript correction of Julius Paul Pradeep John is highly appreciated.

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  1. Department of Biochemistry and Molecular Biology, Capital Medical University, No. 10 Xitoutiao, You An Men Wai, 100069, Beijing, People’s Republic of China

    Junfang Zheng, Hui Shen, Ying Xiong, Xiaomei Yang & Junqi He

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  1. Junfang Zheng
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  2. Hui Shen
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Correspondence to Junqi He.

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J. Zheng and H. Shen contributed equally to this work.

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Zheng, J., Shen, H., Xiong, Y. et al. The β1-adrenergic receptor mediates extracellular signal-regulated kinase activation via Gαs. Amino Acids 38, 75–84 (2010). https://doi.org/10.1007/s00726-008-0207-6

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