scholarly journals Absence of Spontaneous Peroxisome Proliferation in Enoyl-CoA Hydratase/l-3-Hydroxyacyl-CoA Dehydrogenase-deficient Mouse Liver

1999 ◽  
Vol 274 (22) ◽  
pp. 15775-15780 ◽  
Author(s):  
Chao Qi ◽  
Yijun Zhu ◽  
Jie Pan ◽  
Nobuteru Usuda ◽  
Nobuyo Maeda ◽  
...  
Keyword(s):  
Mouse Liver ◽  
Peroxisome Proliferation ◽  
Deficient Mouse ◽  
Hydroxyacyl Coa Dehydrogenase ◽  
Enoyl Coa Hydratase

scholarly journals Induction, immunochemical identity and immunofluorescence localization of an 80000-molecular-weight peroxisome-proliferation-associated polypeptide (polypeptide PPA-80) and peroxisomal enoyl-CoA hydratase of mouse liver and renal cortex

1981 ◽  
Vol 198 (1) ◽  
pp. 177-186 ◽  
Author(s):  
Narendra D. Lalwani ◽  
M. Kumudavalli Reddy ◽  
Mai Mangkornkanok-Mark ◽  
Janardan K. Reddy
Keyword(s):  
Mouse Liver ◽  
Gold Complex ◽  
Kidney Cortex ◽  
Peroxisome Proliferation ◽  
Tubular Epithelium ◽  
Ultrastructural Studies ◽  
Heat Labile ◽  
Immunofluorescence Studies ◽  
Liver And Kidney ◽  
Enoyl Coa Hydratase

The hypolipidaemic drugs methyl clofenapate, BR-931, Wy-14643 and procetofen induced a marked proliferation of peroxisomes in the parenchymal cells of liver and the proximal-convoluted-tubular epithelium of mouse kidney. The proliferation of peroxisomes was associated with 6–12-fold increase in the peroxisomal palmitoyl-CoA oxidizing capacity of the mouse liver. Enhanced activity of the peroxisomal palmitoyl-CoA oxidation system was also found in the renal-cortical homogenates of hypolipidaemic-drug-treated mice. The activity of enoyl-CoA hydratase in the mouse liver increased 30–50-fold and in the kidney cortex 3–5-fold with hypolipidaemic-drug-induced peroxisome proliferation in these tissues, and over 95% of this induced activity was found to be heat-labile peroxisomal enzyme in both organs. Sodium dodecyl sulphate/polyacrylamide-gel-electrophoretic analysis of large-particle and microsomal fractions obtained from the liver and kidney cortex of mice treated with hypolipidaemic peroxisome proliferators demonstrated a substantial increase in the quantity of an 80000-mol.wt. peroxisome-proliferation-associated polypeptide (polypeptide PPA-80). The heat-labile peroxisomal enoyl-CoA hydratase was purified from the livers of mice treated with the hypolipidaemic drug methyl clofenapate; the antibodies raised against this electrophoretically homogeneous protein yielded a single immunoprecipitin band with purified mouse liver enoyl-CoA hydratase and with liver and kidney cortical extracts of normal and hypolipidaemic-drug-treated mice. These anti-(mouse liver enoyl-CoA hydratase) antibodies also cross-reacted with purified rat liver enoyl-CoA hydratase and with the polypeptide PPA-80 obtained from rat and mouse liver. Immunofluorescence studies with anti-(polypeptide PPA-80) and anti-(peroxisomal enoyl-CoA hydratase) provided visual evidence for the localization and induction of polypeptide PPA-80 and peroxisomal enoyl-CoA hydratase in the liver and kidney respectively of normal and hypolipidaemic-drug-treated mice. In the kidney, the distribution of these two proteins is identical and limited exclusively to the cytoplasm of proximal-convoluted-tubular epithelium. The immunofluorescence studies clearly complement the biochemical and ultrastructural observations of peroxisome induction in the liver and kidney cortex of mice fed on hypolipidaemic drugs. In addition, preliminary ultrastructural studies with the protein-A–gold-complex technique demonstrate that the heat-labile hepatic enoyl-CoA hydratase is localized in the peroxisome matrix.

scholarly journals Immunochemical identity of peroxisomal enoyl-CoA hydratase with the peroxisome-proliferation -associated 80,000 mol wt polypeptide in rat liver

1981 ◽  
Vol 89 (3) ◽  
pp. 406-417 ◽  
Author(s):  
MK Reddy ◽  
SA Qreshi ◽  
PF Hollenberg ◽  
JK Reddy
Keyword(s):  
Rat Liver ◽  
Fatty Acyl ◽  
Polyacrylamide Gel ◽  
Lipid Lowering ◽  
Peroxisome Proliferation ◽  
Peroxisome Proliferators ◽  
Heat Labile ◽  
Hydratase Activity ◽  
Oxidation System ◽  
Enoyl Coa Hydratase

Peroxisome proliferators, which induce proliferation of hepatic peroxisomes, have been shown previously to cause a marked increase in an 80,000 mol wt polypeptide predominantly in the light mitochondrial and microsomal fractions of liver of rodents. We now present evidence to show that this hepatic peroxisome-proliferation-associated polypeptide, referred to as polypeptide PPA-80, is immunochemically identical with the multifunctional peroxisome protein displaying heat-labile enoyl-CoA hydratase activity. This conclusion is based on the following observations: (a) the purified polypeptide PPA-80 and the heat- labile enoyl-CoA hydratase from livers of rats treated with the peroxisome proliferators Wy-14,643 {[4-chloro-6(2,3-xylidino)-2-pyrimidinylthio]acetic acid} exhibit identical minimum molecular weights of approximately 80,000 on SDS polyacrylamide gel electrophoresis; (b) these two proteins are immunochemically identical on the basis of ouchterlony double diffusion, immunotitration, rocket immunoelectrophoresis, and crossed immunoelectrophoresis analysis; and (c) the immunoprecipitates formed by antibodies to polypeptide PPA-80 when dissociated on a sephadex G-200 column yield enoyl-CoA hydratase activity. Whether the polypeptide PPA-80 exhibits the activity of other enzyme(s) of the peroxisomal β-oxidation system such as fatty acyl-CoA oxidase activity or displays immunochemical identity with such enzymes remains to be determined. The availability of antibodies to polypeptide PPA-80 and enoyl-CoA hydratase facilitated immunofluorescent and immunocytochemical localization of the polypeptide PPA- 80 and enoyl-CoA hydratase in the rat liver. The indirect immunofluorescent studies with these antibodies provided direct visual evidence for the marked induction of polypeptide PPA-80 and enoyl-CoA hydratase in the livers of rats treated with Wy-14,643. The present studies also provide immunocytochemical evidence for the localization of polypeptide PPA- 80 and the heat-labile enoyl-CoA hydratase in the peroxisome, but not in the mitochondria, of hepatic parenchymal cells. These studies, therefore, provide morphological evidence for the existence of fatty acyl-CoA oxidizing system in peroxisomes. An increase of polypeptide PPA-80 on SDS polyacrylamide gel electrophoretic analysis of the subcellular fractions of liver of rodents treated with lipid-lowering drugs should serve as a reliable and sensitive indicator of enhanced peroxisomal β- oxidation system.

Amino acid and nucleotide sequences of human peroxisomal enoyl-CoA hydratase : 3-hydroxyacyl-CoA dehydrogenase cDNA

1998 ◽  
Vol 21 (1) ◽  
pp. 23-28 ◽  
Author(s):  
S. Fukuda ◽  
Y. Suzuki ◽  
N. Shimozawa ◽  
Z. Zhang ◽  
T. Orii ◽  
...  
Keyword(s):  
Amino Acid ◽  
Nucleotide Sequences ◽  
Hydroxyacyl Coa Dehydrogenase ◽  
Enoyl Coa Hydratase

scholarly journals Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis

1997 ◽  
Vol 328 (2) ◽  
pp. 377-382 ◽  
Author(s):  
Yong-Mei QIN ◽  
M. Antti HAAPALAINEN ◽  
Demara CONRY ◽  
A. Dean CUEBAS ◽  
J. Kalervo HILTUNEN ◽  
...  
Keyword(s):  
Bile Acid ◽  
Acid Synthesis ◽  
Bile Acid Synthesis ◽  
Hydration Reaction ◽  
Multifunctional Enzymes ◽  
Hydratase Activity ◽  
Hydroxyacyl Coa Dehydrogenase ◽  
Liver Peroxisomes ◽  
Enoyl Coa Hydratase

Rat liver peroxisomes contain two multifunctional enzymes: (1) perMFE-1 [2-enoyl-CoA hydratase 1/Δ3,Δ2-enoyl-CoA isomerase/(S)-3-hydroxyacyl-CoA dehydrogenase] and (2) perMFE-2 [2-enoyl-CoA hydratase 2/(R)-3-hydroxyacyl-CoA dehydrogenase]. To investigate the role of the hydratase activity of perMFE-2 in β-oxidation, a truncated version of perMFE-2 was expressed in Escherichia coli as a recombinant protein. The protein catalyses the hydration of straight-chain (2E)-enoyl-CoAs to (3R)-hydroxyacyl-CoAs, but it is devoid of hydratase 1 [(2E)-enoyl-CoA to (3S)-hydroxyacyl-CoA] and (3R)-hydroxyacyl-CoA dehydrogenase activities. The purified enzyme (46 kDa hydratase 2) can be stored as an active enzyme for at least half a year. The recombinant enzyme hydrates (24E)-3α,7α,12α-trihydroxy- 5β-cholest-24-enoyl-CoA to (24R,25R)-3α,7α,12α,24-tetrahydroxy-5β-cholestanoyl-CoA, which has previously been characterized as a physiological intermediate in bile acid synthesis. The stereochemistry of the products indicates that the hydration reaction catalysed by the enzyme proceeds via a syn mechanism. A monofunctional 2-enoyl-CoA hydratase 2 has not been observed as a wild-type protein. The recombinant 46 kDa hydratase 2 described here survives in a purified form under storage, thus being the first protein of this type amenable to application as a tool in metabolic studies.

scholarly journals Identification and characterization of the 2-enoyl-CoA hydratases involved in peroxisomal β-oxidation in rat liver

1997 ◽  
Vol 321 (1) ◽  
pp. 253-259 ◽  
Author(s):  
Martine DIEUAIDE-NOUBHANI ◽  
Dmitry NOVIKOV ◽  
Joël VANDEKERCKHOVE ◽  
Paul P. Van VELDHOVEN ◽  
Guy P. MANNAERTS
Keyword(s):  
Rat Liver ◽  
Hydroxysteroid Dehydrogenase ◽  
Acid Synthesis ◽  
Bile Acid Synthesis ◽  
Side Chain ◽  
Multifunctional Protein ◽  
Hydratase Activity ◽  
Hydroxyacyl Coa Dehydrogenase ◽  
Enoyl Coa Hydratase

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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