Division of Biological Chemistry, Biocenter, Innsbruck Medical University, A-6020 Innsbruck, Austria.
J Lipid Res. 2010 Jun;51(6):1554-9. doi: 10.1194/jlr.D002220. Epub 2009 Nov 5.
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.
脂肪醛脱氢酶（EC 1.2.1.48）将长链脂肪醛转化为相应的酸。这种酶的缺乏会导致 Sjogren Larsson 综合征，这是一种罕见的遗传性疾病，其特征为鱼鳞癣、痉挛和智力迟钝。我们使用荧光醛 pyrenedecanal 和 HPLC 与荧光检测，开发了一种新的方法，通过定量测定产物 pyrenedecanoic 酸以及底物 pyrenedecanal 和可能的副产物，如醛加合物，来监测脂肪醛脱氢酶的活性。如重组酶所示，与其他醛脱氢酶相比，pyrenedecanal 对脂肪醛脱氢酶具有很高的偏好性。该方法允许检测纳克量的微粒体或组织蛋白以及微克量的 Sjogren Larsson 综合征患者皮肤成纤维蛋白中的脂肪醛脱氢酶活性。它可以成功地适应从低温 SDS-PAGE 凝胶切片中分析脂肪醛脱氢酶活性，表明从溶解的大鼠肝微粒体中分离出的脂肪醛脱氢酶活性作为二聚体迁移。因此，通过 HPLC 与荧光检测监测 pyrenedecanal 形成 pyrenedecanoic 酸提供了一种用于测定脂肪醛脱氢酶活性的稳健而敏感的方法。
