Delta 3,5, delta 2,4-dienoyl-CoA isomerase from rat liver mitochondria. Purification and characterization of a new enzyme involved in the beta-oxidation of unsaturated fatty acids.
Mitochondrial delta 3,5, delta 2,4-dienoyl-CoA isomerase, which catalyzes the conversion of 3,5-octadienoyl-CoA to 2,4-octadienoyl-CoA, was purified from rat liver 370-fold at almost 30% yield by a six-step purification procedure. The final preparation appeared to be homogeneous as judged by gel electrophoresis. The molecular weights of the native enzyme and its subunit(s) were estimated to be 126,000 and 32,000, respectively. The purification of delta 3,5, delta 2,4-dienoyl-CoA isomerase completes the characterization of the enzymes functioning in the NADPH-dependent pathway for the beta-oxidation of unsaturated fatty acids with double bonds extending from odd-numbered carbon atoms. This novel pathway may not be operative in peroxisomes because delta 3,5, delta 2,4-dienoyl-CoA isomerase was only detected in mitochondria. Substrates of this pathway are 2,5-dienoyl-CoAs formed from 5-enoyl-CoAs by acyl-CoA dehydrogenase. Two sequential isomerization reactions catalyzed by delta 3, delta 2-enoyl-CoAs isomerase and delta 3,5, delta 2,4-dienoyl-CoA isomerase, respectively, convert 2,5-dienoyl-CoAs to 2,4-dienoyl-CoAs, which are reduced by NADPH-dependent 2,4-dienoyl-CoA reductase (EC 1.3.1.34) before reentering the beta-oxidation spiral.
Luo
M J
MJ
Department of Chemistry, City College, City University of New York, New York 10031.
Smeland
T E
TE
Shoukry
K
K
Schulz
H
H
D000818:Animals; D019748:Carbon-Carbon Double Bond Isomerases; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D005231:Fatty Acids, Unsaturated / Q000302:isolation & purification / Q000378:metabolism; D007535:Isomerases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D008930:Mitochondria, Liver / Q000201:enzymology*; D008954:Models, Biological; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley; D013053:Spectrophotometry; D013347:Subcellular Fractions / Q000201:enzymology
D005231:Fatty Acids, Unsaturated; D046911:Macromolecular Substances; D017886:Durapatite; D007535:Isomerases; D019748:Carbon-Carbon Double Bond Isomerases; C085575:delta(3,5),delta(2,4)-dienoyl-CoA isomerase