The measurement of mitochondrial beta-oxidation by release of 3H2O from [9,10-3H]hexadecanoate: application to skeletal muscle and the use of inhibitors as models of metabolic disease.
We describe the use of a simple assay for beta-oxidation which depends on the release of 3H2O from [9,10-3H]hexadecanoate. This was compared with the use of [1-14C]hexadecanoate which gave comparable results when all the products of beta-oxidation were measured. The prediction that 75% of the tritium is released as 3H2O and 25% as [2-3H]acetyl units was confirmed. The assay was used successfully to demonstrate impaired beta-oxidation in tissue preparations from rats treated with etomoxir and methylenecyclopropylpyruvate which are known inhibitors of beta-oxidation. Abnormalities of beta-oxidation were also detected in skeletal muscle from patients with defects of mitochondrial oxidation.
Kler
R S
RS
Division of Clinical Neuroscience Medical School, University of Newcastle upon Tyne, United Kingdom.
Sherratt
H S
HS
Turnbull
D M
DM
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D002952:Citric Acid Cycle; D004852:Epoxy Compounds / Q000494:pharmacology*; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007005:Hypoglycins / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D008659:Metabolic Diseases / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D008931:Mitochondria, Muscle / Q000187:drug effects / Q000378:metabolism*; D008954:Models, Biological; D010084:Oxidation-Reduction; D010169:Palmitic Acids / Q000378:metabolism*; D011865:Radioisotope Dilution Technique; D051381:Rats; D011919:Rats, Inbred Strains; D014316:Tritium; D014867:Water
D004852:Epoxy Compounds; D007004:Hypoglycemic Agents; D007005:Hypoglycins; D010169:Palmitic Acids; D014867:Water; D014316:Tritium; C022149:ketohypoglycin; C054207:etomoxir