A high-performance liquid chromatographic assay for acid ceramidase activity in cultured fibroblasts from patients with Farber's disease and from controls.
The high-performance liquid chromatographic (HPLC) method we devised for assay of acid ceramidase activity involves coupling of a fluorescent probe to the enzymatically released sphingosine in the reaction mixture and detection of the fluorescent sphingosine derivative by reverse-phase HPLC. Using the method, acid ceramidase activity in fibroblast homogenates was accurately assayed, with or without the addition of exogenous ceramide, as the substrate, and the patients and carriers of Farber's disease could be readily diagnosed.
Mitsuo
K
K
Department of Neurology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Kobayashi
T
T
Shinnoh
N
N
Goto
I
I
D055573:Acid Ceramidase; D000581:Amidohydrolases / Q000032:analysis*; D002478:Cells, Cultured; D055572:Ceramidases; D002851:Chromatography, High Pressure Liquid; D005347:Fibroblasts / Q000201:enzymology; D006579:Heterozygote; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008052:Lipid Metabolism, Inborn Errors / Q000175:diagnosis / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D013110:Sphingosine / Q000032:analysis
D000581:Amidohydrolases; C526693:ASAH1 protein, human; D055573:Acid Ceramidase; D055572:Ceramidases; D013110:Sphingosine