The oxidation of dicarboxylic acid CoA esters via peroxisomal fatty acyl-CoA oxidase.
Evidence supporting a common peroxisomal beta-oxidation pathway for the coenzyme A thioesters of medium-chain-length dicarboxylic acids (DCn-CoA) and monocarboxylic acids (MCn-CoA) has been obtained. Using the mono-CoA esters of dodecanedioic acid (DC12-CoA) and lauroyl-CoA (MC12-CoA) as substrates, parallel inductions of activities and parallel increases in specific activities during purification of peroxisomal fatty acyl-CoA oxidase (EC 1.3.99.3) from rat liver after di(2-ethylhexyl)phthalate treatment were seen. The purified enzyme was used for antiserum production in rabbits; antiserum specificity was verified by immunoblot analysis. Coincident losses of oxidase activities with MC12-CoA and DC12-CoA were found in immunotitration experiments with rat liver homogenates, supporting the hypothesis that peroxisomal fatty acyl-CoA oxidase is solely responsible for the oxidation of medium-chain length dicarboxylic acid substrates. Kinetic studies with purified enzyme using the mono-CoA esters of sebacic (DC10-CoA), suberic (DC8-CoA), and adipic (DC6-CoA) acids along with DC12-CoA revealed substrate inhibition. Although these substrates exhibited similar calculated Vmax values, with decreasing chain length, the combination of increasing Km values and decreasing substrate inhibition constant (Ki) caused the maximum obtainable velocity to decrease. These studies offer an explanation for the previously observed limit of the ability of peroxisomes to chain-shorten dicarboxylates and increased urinary excretion of adipic acid when peroxisomal oxidation of dicarboxylic acids is enhanced.
Poosch
M S
MS
Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201.
Yamazaki
R K
RK
D042962:Acyl-CoA Oxidase; D000818:Animals; D003065:Coenzyme A / Q000378:metabolism*; D003998:Dicarboxylic Acids / Q000378:metabolism*; D004051:Diethylhexyl Phthalate / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D004952:Esters; D007163:Immunosorbent Techniques; D007700:Kinetics; D008099:Liver / Q000201:enzymology* / Q000648:ultrastructure; D008297:Male; D008830:Microbodies / Q000201:enzymology*; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D003998:Dicarboxylic Acids; D004952:Esters; D004051:Diethylhexyl Phthalate; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase; D003065:Coenzyme A