Purification and properties of human D-3-hydroxyacyl-CoA dehydratase: medium-chain enoyl-CoA hydratase is D-3-hydroxyacyl-CoA dehydratase.
Human medium-chain enoyl-CoA hydratase was purified from liver, because we noticed the presence of a high medium-chain enoyl-CoA hydratase activity in human skin fibroblasts catalyzed by an enzyme different from the known enzymes catalyzing the enoyl-CoA hydratase reaction. Two enzyme preparations were obtained. One of them, preparation I, consisted of 46-kDa polypeptide, and its molecular mass was estimated to be 86 kDa. The other, preparation II, consisted of a major 77-kDa polypeptide and minor smaller polypeptides including 46-kDa polypeptide. The molecular mass of preparation II was 154 kDa. Both enzyme preparations catalyzed reversible dehydration of medium-chain D-3-hydroxyacyl-CoA to 2-trans-enoyl-CoA, but did not react with L-3-hydroxyacyl-CoA. Catalytic properties and immunochemical reactivities of these enzyme preparations were nearly the same. The cross-reactive material to the antibody was confirmed to be in peroxisomes by immunohistochemical study of cultured human skin fibroblasts.
Department of Biochemistry, Shinshu University School of Medicine.
L L
LL
Jiang
A
A
Kobayashi
H
H
Matsuura
H
H
Fukushima
T
T
Hashimoto
D002478:Cells, Cultured; D002845:Chromatography; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D006801:Humans; D006836:Hydro-Lyases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology*; D008830:Microbodies / Q000201:enzymology; D008970:Molecular Weight; D012867:Skin / Q000201:enzymology; D013379:Substrate Specificity
D017886:Durapatite; C065185:D-3-hydroxyacyl CoA dehydratase; D006836:Hydro-Lyases