TY - JOUR
T1 - Monitoring of fatty aldehyde dehydrogenase by formation of pyrenedecanoic acid from pyrenedecanal
AU - Keller, Markus A.
AU - Watschinger, Katrin
AU - Golderer, Georg
AU - Maglione, Manuel
AU - Sarg, Bettina
AU - Lindner, Herbert H.
AU - Werner-Felmayer, Gabriele
AU - Terrinoni, Alessandro
AU - Wanders, Ronald J A
AU - Werner, Ernst R.
PY - 2010/6/1
Y1 - 2010/6/1
N2 - 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.
AB - 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.
KW - Dimer
KW - Fatty aldehyde metabolism
KW - Long-chain aldehyde dehydrogenase
KW - Low-temperature SDS gel
KW - Sjogren Larsson syndrome
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U2 - 10.1194/jlr.D002220
DO - 10.1194/jlr.D002220
M3 - Article
C2 - 19965611
AN - SCOPUS:77952698947
VL - 51
SP - 1554
EP - 1559
JO - Journal of Lipid Research
JF - Journal of Lipid Research
SN - 0022-2275
IS - 6
ER -