title: Monitoring of fatty aldehyde dehydrogenase by formation of pyrenedecanoic acid from pyrenedecanal.
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 fluorescence 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 fluorescence detection provides a robust and sensitive method for determination of fatty aldehyde dehydrogenase activity.
authors: Division of Biological Chemistry, Biocenter, Innsbruck Medical University, A-6020 Innsbruck, Austria.
authors: Markus A
authors: MA
authors: Keller
authors: Katrin
authors: K
authors: Watschinger
authors: Georg
authors: G
authors: Golderer
authors: Manuel
authors: M
authors: Maglione
authors: Bettina
authors: B
authors: Sarg
authors: Herbert H
authors: HH
authors: Lindner
authors: Gabriele
authors: G
authors: Werner-Felmayer
authors: Alessandro
authors: A
authors: Terrinoni
authors: Ronald J A
authors: RJ
authors: Wanders
authors: Ernst R
authors: ER
authors: Werner
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D003652:Decanoic Acids / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays / Q000379:methods*; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D009038:Motion; D011721:Pyrenes / Q000378:metabolism*; D051381:Rats; D013696:Temperature
chemical_list: D000447:Aldehydes; D003652:Decanoic Acids; D005456:Fluorescent Dyes; D011721:Pyrenes; C021247:pyrenedecanoic acid; D000445:Aldehyde Oxidoreductases; C038828:long-chain-aldehyde dehydrogenase