Inhibition of dihydroceramide desaturase activity by the sphingosine kinase inhibitor SKI II

J Lipid Res. 2014 Aug;55(8):1711-20. doi: 10.1194/jlr.M049759. Epub 2014 May 29.

Abstract

Sphingosine kinase inhibitor (SKI) II has been reported as a dual inhibitor of sphingosine kinases (SKs) 1 and 2 and has been extensively used to prove the involvement of SKs and sphingosine-1-phosphate (S1P) in cellular processes. Dihydroceramide desaturase (Des1), the last enzyme in the de novo synthesis of ceramide (Cer), regulates the balance between dihydroceramides (dhCers) and Cers. Both SKs and Des1 have interest as therapeutic targets. Here we show that SKI II is a noncompetitive inhibitor (Ki = 0.3 μM) of Des1 activity with effect also in intact cells without modifying Des1 protein levels. Molecular modeling studies support that the SKI II-induced decrease in Des1 activity could result from inhibition of NADH-cytochrome b5 reductase. SKI II, but not the SK1-specific inhibitor PF-543, provoked a remarkable accumulation of dhCers and their metabolites, while both SKI II and PF-543 reduced S1P to almost undetectable levels. SKI II, but not PF543, reduced cell proliferation with accumulation of cells in the G0/G1 phase. SKI II, but not PF543, induced autophagy. These overall findings should be taken into account when using SKI II as a pharmacological tool, as some of the effects attributed to decreased S1P may actually be caused by augmented dhCers and/or their metabolites.

Keywords: autophagy; cell cycle; enzyme inhibition; mass spectrometry; molecular docking; reduced nicotinamide adenine dinucleotide-cytochrome b5 reductase; sphingolipids; sphingosine kinase inhibitor II.

MeSH terms

  • Autophagy / drug effects*
  • Enzyme Inhibitors / pharmacology*
  • G1 Phase / drug effects
  • HeLa Cells
  • Humans
  • Oxidoreductases / antagonists & inhibitors*
  • Oxidoreductases / metabolism*
  • Phosphotransferases (Alcohol Group Acceptor) / antagonists & inhibitors*
  • Pyrrolidines / pharmacology
  • Resting Phase, Cell Cycle / drug effects
  • Sulfones / pharmacology

Substances

  • Enzyme Inhibitors
  • PF-543
  • Pyrrolidines
  • Sulfones
  • Oxidoreductases
  • dihydroceramide desaturase
  • Phosphotransferases (Alcohol Group Acceptor)
  • sphingosine kinase
  • sphingosine kinase 2, human