Monitoring of fatty aldehyde dehydrogenase by formation of pyrenedecanoic acid from pyrenedecanal

Markus A. Keller, Katrin Watschinger, Georg Golderer, Manuel Maglione, Bettina Sarg, Herbert H. Lindner, Gabriele Werner-Felmayer, Alessandro Terrinoni, Ronald J A Wanders, Ernst R. Werner

Research output: Contribution to journalArticlepeer-review

Abstract

Fatty aldehyde dehydrogenase (EC 1.2.1.48) converts long-chain fatty aldehydes to the corresponding acids. Deficiency in this enzyme causes the Sjogren Larsson Syndrome, a rare inherited disorder characterized by ichthyosis, spasticity, and mental retardation. Using a fluorescent aldehyde, pyrenedecanal, and HPLC with fl uorescence detection, we developed a novel method to monitor fatty aldehyde dehydrogenase activity by quantification of the product pyrenedecanoic acid together with the substrate pyrenedecanal and possible side products, such as aldehyde adducts. As shown with recombinant enzymes, pyrenedecanal showed a high preference for fatty aldehyde dehydrogenase compared with other aldehyde dehydrogenases. The method allowed detection of fatty aldehyde dehydrogenase activity in nanogram amounts of microsomal or tissue protein and microgram amounts of Sjogren Larsson syndrome patients' skin fibroblast protein. It could successfully be adapted for the analysis of fatty aldehyde dehydrogenase activity in gel slices derived from low-temperature SDS-PAGE, showing that fatty aldehyde dehydrogenase activity from solubilized rat liver microsomes migrates as a dimer. Thus, monitoring of pyrenedecanoic acid formation from pyrenedecanal by HPLC with fl uorescence detection provides a robust and sensitive method for determination of fatty aldehyde dehydrogenase activity.

Original languageEnglish
Pages (from-to)1554-1559
Number of pages6
JournalJournal of Lipid Research
Volume51
Issue number6
DOIs
Publication statusPublished - Jun 1 2010

Keywords

  • Dimer
  • Fatty aldehyde metabolism
  • Long-chain aldehyde dehydrogenase
  • Low-temperature SDS gel
  • Sjogren Larsson syndrome

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Fingerprint Dive into the research topics of 'Monitoring of fatty aldehyde dehydrogenase by formation of pyrenedecanoic acid from pyrenedecanal'. Together they form a unique fingerprint.

Cite this