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Overexpression of AMP‐metabolizing enzymes controls adenine nucleotide levels and AMPK activation in HEK293T cells
Author(s) -
Plaideau Catheline,
Liu Jianming,
HartleibGeschwindner Judith,
BastinCoyette Laurent,
Bontemps Françoise,
Oscarsson Jan,
Hue Louis,
Rider Mark H.
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.11-198168
Subject(s) - oligomycin , ampk , hek 293 cells , amp deaminase , adenylate kinase , chemistry , transfection , intracellular , enzyme , atpase , microbiology and biotechnology , biochemistry , biology , protein kinase a , adenosine deaminase , gene
We investigated whether overexpression of AMP‐metabolizing enzymes in intact cells would modulate oligomycin‐induced AMPK activation. Human embryonic kidney (HEK) 293T cells were transiently transfected with increasing amounts of plasmid vectors to obtain a graded increase in overexpression of AMP‐deaminase (AMPD) 1, AMPD2, and soluble 5'‐nucleotidase IA (cN‐IA) for measurements of AMPK activation and total intracellular adenine nucleotide levels induced by oligomycin treatment. Overexpression of AMPD1 and AMPD2 slightly decreased AMP levels and oligomycin‐induced AMPK activation. Increased overexpression of cN‐IA led to reductions in the oligomycin‐induced increases in AMP and ADP concentrations by ~70 and 50%, respectively, concomitant with a 50% decrease in AMPK activation. The results support the view that a rise in ADP as well as AMP is important for activation of AMPK, which can thus be regulated by the adenylate energy charge. The control coefficient of cN‐IA on AMP was 0.3‐0.7, whereas the values for AMPD1 and AMPD2 were >0.1, suggesting that in this model cN‐IA exerts a large proportion of control over intracellular AMP. Therefore, small molecule inhibition of cN‐IA could be a strategy for AMPK activation.—Plaideau, C., Liu, J., Hartleib‐Geschwindner, J., Bastin‐Coyette, L., Bontemps, F., Oscarsson, J., Hue, L., Rider, M. H. Overexpression of AMP‐metabolizing enzymes controls adenine nucleotide levels and AMPK activation in HEK293T cells. FASEB J. 26, 2685‐2694 (2012). www.fasebj.org