Inhibition of AMP Kinase by the Protein Phosphatase 2A Heterotrimer, PP2APpp2r2d

J Biol Chem. 2015 Apr 24;290(17):10588-98. doi: 10.1074/jbc.M114.626259. Epub 2015 Feb 18.

Abstract

AMP kinase is a heterotrimeric serine/threonine protein kinase that regulates a number of metabolic processes, including lipid biosynthesis and metabolism. AMP kinase activity is regulated by phosphorylation, and the kinases involved have been uncovered. The particular phosphatases counteracting these kinases remain elusive. Here we discovered that the protein phosphatase 2A heterotrimer, PP2A(Ppp2r2d), regulates the phosphorylation state of AMP kinase by dephosphorylating Thr-172, a residue that activates kinase activity when phosphorylated. Co-immunoprecipitation and co-localization studies indicated that PP2A(Ppp2r2d) directly interacted with AMP kinase. PP2A(Ppp2r2d) dephosphorylated Thr-172 in rat aortic and human vascular smooth muscle cells. A positive correlation existed between decreased phosphorylation, decreased acetyl-CoA carboxylase Acc1 phosphorylation, and sterol response element-binding protein 1c-dependent gene expression. PP2A(Ppp2r2d) protein expression was up-regulated in the aortas of mice fed a high fat diet, and the increased expression correlated with increased blood lipid levels. Finally, we found that the aortas of mice fed a high fat diet had decreased AMP kinase Thr-172 phosphorylation, and contained an Ampk-PP2A(Ppp2r2d) complex. Thus, PP2A(Ppp2r2d) may antagonize the aortic AMP kinase activity necessary for maintaining normal aortic lipid metabolism. Inhibiting PP2A(Ppp2r2d) or activating AMP kinase represents a potential pharmacological treatment for many lipid-related diseases.

Keywords: Cholesterol; Diet; Kinase; Lipid; Phosphatase; Phosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / antagonists & inhibitors*
  • AMP-Activated Protein Kinases / chemistry
  • Animals
  • Aorta / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase / metabolism
  • Cell Line
  • Cholesterol / metabolism
  • Diet, High-Fat / adverse effects
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Lipid Metabolism
  • Lipogenesis
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Okadaic Acid / pharmacology
  • Phosphorylation
  • Protein Phosphatase 2 / antagonists & inhibitors
  • Protein Phosphatase 2 / chemistry*
  • Protein Phosphatase 2 / metabolism*
  • Protein Structure, Quaternary
  • Protein Subunits
  • Protein-Serine-Threonine Kinases / metabolism
  • RNA, Small Interfering / genetics
  • Rats
  • Sterol Regulatory Element Binding Protein 1 / metabolism

Substances

  • Enzyme Inhibitors
  • Protein Subunits
  • RNA, Small Interfering
  • Srebf1 protein, rat
  • Sterol Regulatory Element Binding Protein 1
  • Okadaic Acid
  • Cholesterol
  • Protein-Serine-Threonine Kinases
  • Stk11 protein, rat
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase
  • AMP-Activated Protein Kinases
  • Protein Phosphatase 2