A novel method for determining peroxisomal fatty acid β-oxidation.
The purpose of this study is to establish an assay method to screen for chemical compounds that stimulate peroxisomal fatty acid β-oxidation activity in X-linked adrenoleukodystropy (X-ALD) fibroblasts. In this investigation, we used 12-(1-pyrene)dodecanoic acid (pyrene-C12:0), a fluorescent fatty acid analog, as a substrate for fatty acid β-oxidation. When human skin fibroblasts were incubated with pyrene-C12:0, β-oxidation products such as pyrene-C10:0 and pyrene-C8:0 were generated time-dependently. These β-oxidation products were scarcely detected in the fibroblasts from patients with Zellweger syndrome, a peroxisomal biogenesis disorder. In contrast, in fibroblasts with mitochondrial carnitine-acylcarnitine translocase deficiency, the β-oxidation products were detected at a level similar to control fibroblasts. These results indicate that the β-oxidation of pyrene-C12:0 takes place in peroxisomes, but not mitochondria, so pyrene-C12:0 is useful for measuring peroxisomal fatty acid β-oxidation activity. In X-ALD fibroblasts, the β-oxidation activity for pyrene-C12:0 was approximately 40 % of control fibroblasts, which is consistent with previous results using [1-(14)C]lignoceric acid as the substrate. The present study provides a convenient procedure for screening chemical compounds that stimulate the peroxisomal fatty acid β-oxidation in X-ALD fibroblasts.
Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan. masa@pha.u-toyama.ac.jp.
Masashi
0000-0003-2333-2190
M
Morita
Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Shun
S
Matsumoto
Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Airi
A
Okazaki
Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Kaito
K
Tomita
Institute of Natural Medicine, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Shiro
S
Watanabe
Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Kosuke
K
Kawaguchi
Laboratory of Synthetic and Medicinal Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Daishiro
D
Minato
Laboratory of Synthetic and Medicinal Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Yuji
Y
Matsuya
Division of Genomic Research, Life Science Research Center, Gifu University, Gifu, 501-1193, Japan.
Nobuyuki
N
Shimozawa
Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
Tsuneo
T
Imanaka
D000326:Adrenoleukodystrophy / Q000378:metabolism; D002333:Carnitine Acyltransferases / Q000172:deficiency / Q000378:metabolism; D002478:Cells, Cultured; D005227:Fatty Acids / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007850:Lauric Acids / Q000378:metabolism; D008052:Lipid Metabolism, Inborn Errors / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D010084:Oxidation-Reduction; D018901:Peroxisomal Disorders / Q000378:metabolism; D020675:Peroxisomes / Q000378:metabolism*; D011721:Pyrenes / Q000378:metabolism; D012867:Skin / Q000378:metabolism; D015211:Zellweger Syndrome / Q000378:metabolism
D005227:Fatty Acids; D007850:Lauric Acids; D011721:Pyrenes; C040596:12-(1-pyrene)dodecanoic acid; C030984:pyrene; D002333:Carnitine Acyltransferases; C010210:lignoceric acid