A novel method for determining peroxisomal fatty acid β-oxidation

• Published in: Journal of inherited metabolic disease 2016-09, Vol.39 (5), p.725-731
• Main Authors: Morita, Masashi, Matsumoto, Shun, Okazaki, Airi, Tomita, Kaito, Watanabe, Shiro, Kawaguchi, Kosuke, Minato, Daishiro, Matsuya, Yuji, Shimozawa, Nobuyuki, Imanaka, Tsuneo
• Format: Article
• Language: English
• Subjects: Adrenoleukodystrophy - metabolism; Biochemistry; Carnitine Acyltransferases - deficiency; Carnitine Acyltransferases - metabolism; Cells, Cultured; Fatty Acids - metabolism; Fibroblasts - metabolism; Human Genetics; Humans; Internal Medicine; Lauric Acids - metabolism; Lipid Metabolism, Inborn Errors - metabolism; Medicine; Medicine & Public Health; Metabolic Diseases; Mitochondria - metabolism; Original Article; Oxidation-Reduction; Pediatrics; Peroxisomal Disorders - metabolism; Peroxisomes - metabolism; Pyrenes - metabolism; Skin - metabolism; Zellweger Syndrome - metabolism
• ISSN: 0141-8955; 1573-2665
• DOI: 10.1007/s10545-016-9952-y

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.