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Enzymatic assay for 3-hydroxyacyl-CoA and 2-trans-enoyl-CoA intermediates of beta-oxidation. 1988

P M Latipää, and I E Hassinen, and J K Hiltunen
Department of Medical Biochemistry, University of Oulu, Finland.

An enzymatic assay is presented for determining 3-hydroxyacyl-CoA and 2-trans-enoyl-CoA esters in tissue samples. The procedure includes extraction of acyl-CoA esters from frozen tissue samples with chloroform/methanol, stochiometric oxidation of the acyl esters to 3-keto-acyl-CoAs with 3-hydroxyacyl-CoA dehydrogenase in the presence or absence of enoyl-CoA hydratase, and an enzymatic cycling amplification of NADH produced for fluorometric determination. The procedure allows measurement of these intermediates of beta-oxidation at the picomole level. The method has been used successfully to measure the concentrations of 3-hydroxyacyl-CoA and 2-trans-enoyl-CoA esters in isolated rat hearts perfused with glucose or oleate or under anoxia.

UI MeSH Term Description Entries
D008929 Mitochondria, Heart The mitochondria of the myocardium. Heart Mitochondria,Myocardial Mitochondria,Mitochondrion, Heart,Heart Mitochondrion,Mitochondria, Myocardial
D009243 NAD A coenzyme composed of ribosylnicotinamide 5'-diphosphate coupled to adenosine 5'-phosphate by pyrophosphate linkage. It is found widely in nature and is involved in numerous enzymatic reactions in which it serves as an electron carrier by being alternately oxidized (NAD+) and reduced (NADH). (Dorland, 27th ed) Coenzyme I,DPN,Diphosphopyridine Nucleotide,Nadide,Nicotinamide-Adenine Dinucleotide,Dihydronicotinamide Adenine Dinucleotide,NADH,Adenine Dinucleotide, Dihydronicotinamide,Dinucleotide, Dihydronicotinamide Adenine,Dinucleotide, Nicotinamide-Adenine,Nicotinamide Adenine Dinucleotide,Nucleotide, Diphosphopyridine
D010084 Oxidation-Reduction A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471). Redox,Oxidation Reduction
D011919 Rats, Inbred Strains Genetically identical individuals developed from brother and sister matings which have been carried out for twenty or more generations or by parent x offspring matings carried out with certain restrictions. This also includes animals with a long history of closed colony breeding. August Rats,Inbred Rat Strains,Inbred Strain of Rat,Inbred Strain of Rats,Inbred Strains of Rats,Rat, Inbred Strain,August Rat,Inbred Rat Strain,Inbred Strain Rat,Inbred Strain Rats,Inbred Strains Rat,Inbred Strains Rats,Rat Inbred Strain,Rat Inbred Strains,Rat Strain, Inbred,Rat Strains, Inbred,Rat, August,Rat, Inbred Strains,Rats Inbred Strain,Rats Inbred Strains,Rats, August,Rats, Inbred Strain,Strain Rat, Inbred,Strain Rats, Inbred,Strain, Inbred Rat,Strains, Inbred Rat
D002384 Catalysis The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction. Catalyses
D004746 Enoyl-CoA Hydratase An enzyme that catalyzes reversibly the hydration of unsaturated fatty acyl-CoA to yield beta-hydroxyacyl-CoA. It plays a role in the oxidation of fatty acids and in mitochondrial fatty acid synthesis, has broad specificity, and is most active with crotonyl-CoA. EC 4.2.1.17. 3-Hydroxyacyl CoA Hydrolyases,3-Hydroxyacyl Dehydratases,Crotonase,Enoyl Hydrase,beta-Hydroxyacyl Dehydratases,Enoyl CoA Hydratases,beta-Hydroxyacyl-CoA Dehydrases,trans-2-Enoyl-Coenzyme A Hydratase,3 Hydroxyacyl CoA Hydrolyases,3 Hydroxyacyl Dehydratases,CoA Hydratases, Enoyl,CoA Hydrolyases, 3-Hydroxyacyl,Dehydrases, beta-Hydroxyacyl-CoA,Dehydratases, 3-Hydroxyacyl,Dehydratases, beta-Hydroxyacyl,Enoyl CoA Hydratase,Hydrase, Enoyl,Hydratase, Enoyl-CoA,Hydratase, trans-2-Enoyl-Coenzyme A,Hydratases, Enoyl CoA,Hydrolyases, 3-Hydroxyacyl CoA,beta Hydroxyacyl CoA Dehydrases,beta Hydroxyacyl Dehydratases,trans 2 Enoyl Coenzyme A Hydratase
D005227 Fatty Acids Organic, monobasic acids derived from hydrocarbons by the equivalent of oxidation of a methyl group to an alcohol, aldehyde, and then acid. Fatty acids are saturated and unsaturated (FATTY ACIDS, UNSATURATED). (Grant & Hackh's Chemical Dictionary, 5th ed) Aliphatic Acid,Esterified Fatty Acid,Fatty Acid,Fatty Acids, Esterified,Fatty Acids, Saturated,Saturated Fatty Acid,Aliphatic Acids,Acid, Aliphatic,Acid, Esterified Fatty,Acid, Saturated Fatty,Esterified Fatty Acids,Fatty Acid, Esterified,Fatty Acid, Saturated,Saturated Fatty Acids
D005260 Female Females
D006836 Hydro-Lyases Enzymes that catalyze the breakage of a carbon-oxygen bond leading to unsaturated products via the removal of water. EC 4.2.1. Dehydratase,Dehydratases,Hydrase,Hydrases,Hydro Lyase,Hydro-Lyase,Hydro Lyases,Lyase, Hydro,Lyases, Hydro
D000214 Acyl Coenzyme A S-Acyl coenzyme A. Fatty acid coenzyme A derivatives that are involved in the biosynthesis and oxidation of fatty acids as well as in ceramide formation. Acyl CoA,Fatty Acyl CoA,Long-Chain Acyl CoA,Acyl CoA, Fatty,Acyl CoA, Long-Chain,CoA, Acyl,CoA, Fatty Acyl,CoA, Long-Chain Acyl,Coenzyme A, Acyl,Long Chain Acyl CoA

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