Measuring Intracellular Nucleotides and Changes in AMPK Phosphorylation in Response to Small Molecules — ASN Events
  1. Schedule
  2. Session 10: Poster Session C
  3. Measuring Intracellular Nucleotides and Changes in AMPK Phosphorylation in Response to Small Molecules

Measuring Intracellular Nucleotides and Changes in AMPK Phosphorylation in Response to Small Molecules (#367)

Toby Dite 1 , Chris G Langendorf 1 , Naomi Ling 1 , Zhiping Chen 1 , Sandra Galic 1 , Samah Issa 1 , Matthew T O'Brien 1 , Jon S Oakhill 1 , John W Scott 1 , Bruce E Kemp 1
  1. St Vincent's Institute of Medical Research, Fitzroy, VIC, Australia

The AMP-activated protein kinase (AMPK) is central in regulating energy consumption in cells in response to changing energy availability. Depletion of intracellular adenylate energy charge increases phosphorylation of the α subunit activation loop Thr172 by upstream kinases LKB1 or CaMKKβ1 . In return, AMPK adjusts metabolic pathways, such as fatty acid oxidation and glucose uptake to restore ATP levels in the cell. Direct activators of AMPK, such as A-769662 are therefore being investigated as potential therapeutics for diseases related to metabolism such as obesity and type 2 diabetes2 . A-769662 has been shown to activate AMPK pathways in the cell, however the degree to which this is caused by disruption to mitochondrial energy production has not been shown. Using mass spectrometry, changes in intracellular nucleotide levels in response to changing doses of small molecules have been profiled, as well as corresponding changes to phosphorylation states of AMPK.

  1. Oakhill, J. S., Scott, J. W. & Kemp, B. E. AMPK functions as an adenylate charge-regulated protein kinase. Trends in Endocrinol & Metabolism 23, 125-132 (2012)
  2. Cool, B. et al. Identification and characterization of a small molecule AMPK activator that treats key components of type 2 diabetes and the metabolic syndrome. Cell metabolism 3, 403-416 (2006)