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The metabolic fluxes of cis-5-enoyl-CoAs through the beta-oxidation cycle were studied in solubilized rat liver mitochondrial samples and compared with saturated acyl-CoAs of equal chain length. These studies were accomplished using either spectrophotometric assay of enzyme activities and/or the analysis of metabolites and precursors using a gas chromatographic method after conversion of CoA esters into their free acids. Cis-5-enoyl-CoAs were dehydrogenated by acyl-CoA oxidase or acyl-CoA dehydrogenases at significantly lower rates (10-44%) than saturated acyl-CoAs. However, enoyl-CoA hydratase hydrated trans-2-cis-5-enoyl-CoA at a faster rate (at least 1.5-fold) than trans-2-enoyl-CoA. The combined activities of 3-hydroxyacyl-CoA dehydrogenase and 3-ketoacyl-CoA thiolase for 3-hydroxy-cis-5-enoyl-CoAs derived from cis-5-enoyl-CoAs were less than 40% of the activity for the corresponding 3-hydroxyacyl-CoAs prepared from saturated acyl-CoAs. Rat liver mitochondrial beta-oxidation enzymes were capable of metabolizing cis-5-enoyl-CoA via one cycle of beta-oxidation to cis-3-enoyl-CoA with two less carbons. However, the overall rates of one cycle of beta-oxidation, as determined with stable-isotope-labelled tracer, was only 15-25%, for cis-5-enoyl-CoA, of that for saturated acyl-CoA. In the presence of NADPH, the metabolism of cis-5-enoyl-CoAs was switched to the reduction pathway.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of metabolic fluxes of cis-5-enoyl-CoA and saturated acyl-CoA through the β-oxidation pathway