Peroxisomal beta-oxidation system of Candida tropicalis. Purification of a multifunctional protein possessing enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase and 3-hydroxyacyl-CoA epimerase activities

In this study we attempted to determine the number of 2-enoyl-CoA hydratases involved in peroxisomal β-oxidation. We therefore separated peroxisomal proteins from rat liver on several chromatographic columns and measured hydratase activities on the eluates with different substrates. The results indicate that rat liver peroxisomes contain two hydratase activities: (1) a hydratase activity associated with multifunctional protein 1 (MFP-1) (2-enoyl-CoA hydratase/Δ3,Δ2-enoyl-CoA isomerase/l-3-hydroxyacyl-CoA dehydrogenase) and (2) a hydratase activity associated with MFP-2 (17β-hydroxysteroid dehydrogenase/d-3-hydroxyacyl-CoA dehydrogenase/2-enoyl-CoA hydratase). MFP-1 forms and dehydrogenates l-3-hydroxyacyl-CoA species, whereas MFP-2 forms and dehydrogenates d-3-hydroxyacyl-CoA species. A portion of MFP-2 is proteolytically cleaved, most probably in the peroxisome, into a 34 kDa 17β-hydroxysteroid dehydrogenase/d-3-hydroxyacyl-CoA dehydrogenase and a 45 kDa d-specific 2-enoyl-CoA hydratase. Finally, the results confirm that MFP-1 is involved in the degradation of straight-chain fatty acids, whereas MFP-2 and its cleavage products seem to be involved in the degradation of the side chain of cholesterol (bile acid synthesis)

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Relationship between enoyl-CoA hydratase and a peroxisomal bifunctional enzyme, enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase, from an n-alkane-utilizing yeast, Candida tropicalis.

Agricultural and Biological Chemistry ◽

10.1271/bbb1961.51.2197 ◽

1987 ◽

Vol 51 (8) ◽

pp. 2197-2205 ◽

Cited By ~ 5

Author(s):

Mitsuyoshi UEDA ◽

Tadashi MORHCAWA ◽

Hirofumi OKADA ◽

Atsuo TANAKA

Keyword(s):

Candida Tropicalis ◽

Bifunctional Enzyme ◽

Hydroxyacyl Coa Dehydrogenase ◽

Enoyl Coa Hydratase

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Relationship between Enoyl-CoA Hydratase and a Peroxisomal Bifunctional Enzyme, Enoyl-CoA Hydratase/3-Hydroxyacyl- CoA Dehydrogenase, from ann-Alkane-utilizing Yeast,Candida tropicalis

Agricultural and Biological Chemistry ◽

10.1080/00021369.1987.10868375 ◽

1987 ◽

Vol 51 (8) ◽

pp. 2197-2205

Author(s):

Mitsuyoshi Ueda ◽

Tadashi Morikawa ◽

Hirofumi Okada ◽

Atsuo Tanaka

Keyword(s):

Candida Tropicalis ◽

Bifunctional Enzyme ◽

Hydroxyacyl Coa Dehydrogenase ◽

Enoyl Coa Hydratase

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Acyl-Coa oxidase and hydratase-dehydrogenase, two enzymes of the peroxisomal beta-oxidation system, are synthesized on free polysomes of clofibrate-treated rat liver.

The Journal of Cell Biology ◽

10.1083/jcb.99.6.2241 ◽

1984 ◽

Vol 99 (6) ◽

pp. 2241-2246 ◽

Cited By ~ 58

Author(s):

R A Rachubinski ◽

Y Fujiki ◽

R M Mortensen ◽

P B Lazarow

Keyword(s):

Rat Liver ◽

Translational Efficiency ◽

Beta Oxidation ◽

Sds Page ◽

Reticulocyte Lysate ◽

Normal Rat ◽

Membrane Bound ◽

Hydroxyacyl Coa Dehydrogenase ◽

Acyl Coa Oxidase ◽

Oxidation System

We investigated the site of synthesis of two abundant proteins in clofibrate-induced rat hepatic peroxisomes. RNA was extracted from free and membrane-bound polysomes, heated to improve translational efficiency, and translated in the mRNA-dependent, reticulocyte-lysate-cell-free, protein-synthesizing system. The peroxisomal acyl-CoA oxidase and enoyl-CoA hydratase-beta-hydroxyacyl-CoA dehydrogenase 35S-translation products were isolated immunochemically, analyzed by SDS PAGE and fluorography, and quantitated by densitometric scanning. The RNAs coding for these two peroxisomal proteins were found predominantly on free polysomes, and the translation products co-migrated with the mature proteins. As in normal rat liver, preproalbumin and catalase were synthesized mainly by membrane-bound and by free polysomes, respectively. mRNAs for a number of minor 35S-translation products also retained by the anti-peroxisomal immunoadsorbent were similarly found on free polysomes. These results, together with previous data, allow the generalization that the content proteins of rat liver peroxisomes are synthesized on free polysomes, and the data imply a posttranslational packaging mechanism for these major content proteins.

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