A sensitive spectrophotometric assay for peroxisomal acyl-CoA oxidase.

A simple spectrophotometric assay was developed for peroxisomal fatty acyl-CoA oxidase activity. The assay, based on the H2O2-dependent oxidation of leuco-dichlorofluorescein catalysed by exogenous peroxidase, is more sensitive than methods previously described. By using mouse liver samples, cofactor requirements were assessed and a linear relationship was demonstrated between dye oxidation and enzyme concentration. By using this assay on subcellular fractions, palmitoyl-CoA oxidase activity was localized for the first time in microperoxisomes of rat intestine. The assay was also adapted to measure D-amino acid oxidase activity, demonstrating the versatility of this method for measuring activity of other H2O2-producing oxidases.

[1]  E. Harrison,et al.  Fluorometric assay for rat liver peroxisomal fatty acyl-coenzyme A oxidase activity. , 1983, Journal of lipid research.

[2]  M. Connock,et al.  Detection of acyl-CoA beta-oxidation enzymes in peroxisomes (microperoxisomes) of mouse heart. , 1983, Biochemistry international.

[3]  M. Le Hir,et al.  Peroxisomal and mitochondrial beta-oxidation in the rat kidney: distribution of fatty acyl-coenzyme A oxidase and 3-hydroxyacyl-coenzyme A dehydrogenase activities along the nephron. , 1982, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[4]  M. Connock,et al.  Enhancement by dietary clofibrate of peroxisomal palmityl-CoA oxidase in kidney and small intestine of albino mice and liver of genetically lean and obese mice. , 1981, Life sciences.

[5]  M. Connock,et al.  Palmityl-CoA oxidase: detection in several guinea pig tissues and peroxisomal localisation in mucosa, of small intestine. , 1980, Life sciences.

[6]  N. Inestrosa,et al.  Detection of peroxisomal fatty acyl-coenzyme A oxidase activity. , 1979, The Biochemical journal.

[7]  N. Inestrosa,et al.  Fatty acid oxidation by human liver peroxisomes. , 1979, Biochemical and biophysical research communications.

[8]  J. F. Hogg,et al.  Chain length specificities of peroxisomal and mitochondrial beta-oxidation in rat liver. , 1979, Biochemical and biophysical research communications.

[9]  P. Murphy,et al.  Enzyme activities of isolated hepatic peroxisomes from genetically lean and obese male mice. , 1979, Archives of biochemistry and biophysics.

[10]  B. Gmeiner,et al.  β-oxidation in peroxisomes of brown adipose tissue , 1978 .

[11]  P. Lazarow.  Rat liver peroxisomes catalyze the beta oxidation of fatty acids. , 1978, The Journal of biological chemistry.

[12]  J. von Wartburg,et al.  A sensitive photometric assay for monoamine oxidase. , 1978, Analytical biochemistry.

[13]  H. Shio,et al.  Analytical subcellular fractionation studies on rat liver and on isolated jejunal enterocytes with special reference to the separation of lysosomes, peroxisomes and mitochondria. , 1976, Clinical science and molecular medicine.

[14]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.