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Metformin interacts with AMPK through binding to γ subunit

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Abstract

Metformin acts as an energy regulator by activating 5′-adenosine monophosphate-activated protein kinase (AMPK), which is a key player in the regulation of energy homeostasis, but it is uncertain whether AMPK is its direct target. This study aims to investigate the possible interaction between metformin and AMPK. First, we verified that metformin can promote AMPK activation and induce ACC inactivation in human HepG2 cells using western blot. Then we predicted that metformin may interact with the γ subunit of AMPK by molecular docking analysis. The fluorescence spectrum and ForteBio assays indicated that metformin has a stronger binding ability to the γ subunit of AMPK than to α subunit. In addition, interaction of metformin with γ-AMPK resulted in a decrease in the α-helicity determined by CD spectra, but relatively little change was seen with α-AMPK. These results demonstrate that metformin may interact with AMPK through binding to the γ subunit.

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Abbreviations

AICAR:

5-Aminoimidazole-4-carboxamide-1-β-d-ribonucleoside

ACC:

Acetyl-CoA carboxylase

AMPK:

5′-Adenosine monophosphate-activated protein kinase

BSA:

Bovine serum albumin

CD:

Circular dichroism

DMEM:

Dulbecco’s modified Eagle’s medium

DTT:

Dithiothreitol

ECL:

Enhanced chemiluminescence

EDTA:

Ethylenediaminetetraacetic acid

EGTA:

Ethylene glycol tetraacetic acid

FCS:

Fetal calf serum

IPTG:

Isopropyl-1-thio-d-galactopyranoside

PMSF:

Phenylmethanesulfonyl fluoride

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

ZMP:

5-Aminoimidazole-4-carboxamide-1-β-d-ribotide

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Acknowledgments

We thank the members of the M.P.E. laboratories of Xiamen University and School of Life Sciences of Tsinghua University for providing technology support. This study was supported by National Institutes of Major Research Projects (No. 200909040785) and the Science and Technology Foundation of Fujian Province of China (NO.2010D013).

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Oncology, The 174th Hospital of the Chinese People’s Liberation Army, The Affiliated Chenggong Hospital of Xiamen University, No. 94–96 Wenyuan Road, Xiamen, 361003, Fujian, People’s Republic of China

    Yaya Zhang, Yongjun Wang, Chuanen Bao, Yingyi Xu & Yuqiang Chen

  2. School of Life Sciences, Xiamen University, No. 422 Si Ming South Road, Xiamen, 361005, Fujian, People’s Republic of China

    Huili Shen, Junjie Chen & Jianghua Yan

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  1. Yaya Zhang
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  2. Yongjun Wang
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  5. Huili Shen
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  8. Yuqiang Chen
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Corresponding author

Correspondence to Yuqiang Chen.

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Yaya Zhang and Yongjun Wang contributed equally to this study.

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Zhang, Y., Wang, Y., Bao, C. et al. Metformin interacts with AMPK through binding to γ subunit. Mol Cell Biochem 368, 69–76 (2012). https://doi.org/10.1007/s11010-012-1344-5

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