Epimerization of 3-hydroxyacyl-CoA esters in rat liver. Involvement of two 2-enoyl-CoA hydratases.
Interconversion of D- and L-isomers of 3-hydroxy-decanoyl-CoA was catalyzed by rat liver homogenate. Cation exchange chromatography followed by ammonium sulfate precipitation and PBE-94 chromatofocusing column was used to separate the peroxisomal bifunctional protein, the classic 2-enoyl-CoA hydratase (crotonase), and a novel 2-enoyl-CoA hydratase. Epimerization activity was lost during the last purification step. None of the above proteins was capable of catalyzing the epimerization by itself, but reconstitution was achieved by recombining crotonase and the novel 2-enoyl-CoA hydratase. Since hydration by the latter enzyme follows a different stereochemical course from that with crotonase, these two hydratases are distinguished as 2-enoyl-CoA hydratase 1 (crotonase) and 2-enoyl-CoA hydratase 2 (the novel hydratase). The data strongly suggested that epimerization in the rat liver proceeds via dehydration-hydration catalyzed by the two different hydratases. The intermediate of this two step mechanism appears to be trans-2-enoyl-CoA.
Hiltunen
J K
JK
Abteilung für Zellbiochemie, Medizinische Fakultät, Ruhr-Universität Bochum, Federal Republic of Germany.
Palosaari
P M
PM
Kunau
W H
WH
D000214:Acyl Coenzyme A / Q000378:metabolism*; D000818:Animals; D004746:Enoyl-CoA Hydratase / Q000302:isolation & purification / Q000378:metabolism*; D006836:Hydro-Lyases / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007536:Isomerism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D019998:Racemases and Epimerases / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D000214:Acyl Coenzyme A; D007527:Isoenzymes; C060624:3-hydroxydecanoyl-coenzyme A; D006836:Hydro-Lyases; D004746:Enoyl-CoA Hydratase; D019998:Racemases and Epimerases