Measurement of short-chain acyl-CoA dehydrogenase (SCAD) in cultured skin fibroblasts with hexanoyl-CoA as a competitive inhibitor to eliminate the contribution of medium-chain acyl-CoA dehydrogenase
| Autor: | Adrian C. Sewell, Ronald J.A. Wanders, H. S. A. Heymans, K. E. Niezen-Koning, Gijs T. Nagel |
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| Přispěvatelé: | Other departments, Center for Liver, Digestive and Metabolic Diseases (CLDM) |
| Rok vydání: | 1994 |
| Předmět: |
FATTY-ACID OXIDATION
LIVER Clinical Biochemistry SECONDARY CARNITINE DEFICIENCY Dehydrogenase PATIENT Biochemistry Binding Competitive Sensitivity and Specificity Acyl-CoA Dehydrogenase Gas Chromatography-Mass Spectrometry GAS CHROMATOGRAPHY MASS SPECTROMETRY Substrate Specificity MYOPATHY Acyl-CoA Dehydrogenases medicine Humans FATTY ACID OXIDATION Fibroblast Beta oxidation SHORT-CHAIN ACYL-COA DEHYDROGENASE Cells Cultured Skin chemistry.chemical_classification PURIFICATION Butyryl-CoA dehydrogenase biology Biochemistry (medical) COENZYME-A DEHYDROGENASE Substrate (chemistry) Infant General Medicine Fibroblasts medicine.anatomical_structure Enzyme chemistry DEFECT Enzyme inhibitor INBORN ERROR biology.protein Acyl Coenzyme A Scad |
| Zdroj: | University of Groningen Clinica chimica acta; international journal of clinical chemistry, 229(1-2), 99-106. Elsevier Clinica chimica acta, 229(1-2), 99-106. ELSEVIER SCIENCE BV |
| ISSN: | 0009-8981 |
| Popis: | Short-chain acyl-CoA dehydrogenase (SCAD) deficiency has so far been reported in only very few patients. This is due, in part, to the problems involved in measuring the activity of SCAD unequivocally. The main reason for this difficulty is that butyryl-CoA, the substrate preferably used for SCAD activity measurements, is also dehydrogenated by medium-chain acyl-CoA dehydrogenase (MCAD). Elimination of this contribution can be achieved by means of immune precipitation with a specific MCAD antibody. We now describe a relatively straightforward assay based on the use of gas chromatography/mass spectrometry for detection. The contribution of MCAD to overall butyryl-CoA dehydrogenation was eliminated by adding excess hexanoyl-CoA to the assay medium. The validity of the method developed was checked by SCAD-activity measurements in fibroblasts from an established SCAD-deficient patient. |
| Databáze: | OpenAIRE |
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