Immunoaffinity purification and characterization of mitochondrial membrane-bound D-3-hydroxybutyrate dehydrogenase from Jaculus orientalis.

6533b7d1fe1ef96bd125c235

RESEARCH PRODUCT

Immunoaffinity purification and characterization of mitochondrial membrane-bound D-3-hydroxybutyrate dehydrogenase from Jaculus orientalis.

Mustapha Cherkaoui-malki Pierre Andreoletti Driss Mountassif Adnane Moutaouakkil Zakaria El Kebbaj M'hammed Saïd El Kebbaj Norbert Latruffe

subject

lcsh:Animal biochemistry MESH : Aged MESH : Rodentia MESH: Rodentia MESH: Base Sequence Biochemistry MESH: Lipid Peroxidation MESH : Information Services Antigen-Antibody Reactions MESH: Health Education Epitopes MESH: Organizations MESH: Libraries MESH: Antigen-Antibody Reactions lcsh:QD415-436 MESH: Animals MESH : Organizations MESH : Physician's Role MESH: Bacterial Proteins Immunosorbent Techniques chemistry.chemical_classification MESH: Conserved Sequence Methodology Article MESH : Computer Communication Networks MESH: Chromatography Affinity MESH : Pseudomonas aeruginosa MESH : Chromatography Affinity MESH : Immunosorbent Techniques MESH: Ethnic Groups MESH : Ethnic Groups MESH: Epitopes MESH : Patient Satisfaction MESH : United States MESH: Mitochondria MESH : Antigen-Antibody Reactions Molecular Sequence Data MESH : Hydroxybutyrate Dehydrogenase MESH: Sequence Alignment Rodentia MESH: Information Services MESH : Epitopes lcsh:Biochemistry MESH : Mitochondrial Membranes Bacterial Proteins MESH : Conserved Sequence Complementary DNA MESH : Libraries Molecular Biology [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry Molecular Biology MESH: Immunosorbent Techniques MESH: Molecular Sequence Data MESH: Humans MESH : Consumer Participation MESH : Humans MESH: Adult MESH: Patient Satisfaction MESH: Hydroxybutyrate Dehydrogenase MESH: Consumer Participation chemistry Lipid Peroxidation MESH: Female MESH: Liver MESH : Sequence Analysis DNA MESH: Continental Population Groups MESH: Sequence Analysis DNA MESH : Molecular Sequence Data Dehydrogenase Chromatography Affinity MESH: Mitochondrial Membranes MESH: Antibodies Bacterial MESH : Bacterial Proteins MESH : Female MESH: Computer Communication Networks Conserved Sequence MESH: Aged biology MESH : Lipid Peroxidation MESH : Sequence Alignment MESH: Physician's Role MESH : Adult Antibodies Bacterial Mitochondria Amino acid Liver Biochemistry Mitochondrial Membranes Pseudomonas aeruginosa MESH: Pseudomonas aeruginosa MESH : Mitochondria MESH : Mass Media MESH: Mass Media MESH : Male Hydroxybutyrate Dehydrogenase Affinity chromatography MESH : Health Education [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology MESH: United States Animals [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology MESH : Antibodies Bacterial lcsh:QP501-801 Jaculus orientalis MESH : Continental Population Groups Base Sequence MESH : Liver Sequence Analysis DNA biology.organism_classification Molecular biology MESH: Male Enzyme Polyclonal antibodies biology.protein MESH : Base Sequence NAD+ kinase MESH : Animals Sequence Alignment

description

Abstract Background The interconversion of two important energy metabolites, 3-hydroxybutyrate and acetoacetate (the major ketone bodies), is catalyzed by D-3-hydroxybutyrate dehydrogenase (BDH1: EC 1.1.1.30), a NAD+-dependent enzyme. The eukaryotic enzyme is bound to the mitochondrial inner membrane and harbors a unique lecithin-dependent activity. Here, we report an advanced purification method of the mammalian BDH applied to the liver enzyme from jerboa (Jaculus orientalis), a hibernating rodent adapted to extreme diet and environmental conditions. Results Purifying BDH from jerboa liver overcomes its low specific activity in mitochondria for further biochemical characterization of the enzyme. This new procedure is based on the use of polyclonal antibodies raised against BDH from bacterial Pseudomonas aeruginosa. This study improves the procedure for purification of both soluble microbial and mammalian membrane-bound BDH. Even though the Jaculus orientalis genome has not yet been sequenced, for the first time a D-3-hydroxybutyrate dehydrogenase cDNA from jerboa was cloned and sequenced. Conclusion This study applies immunoaffinity chromatography to purify BDH, the membrane-bound and lipid-dependent enzyme, as a 31 kDa single polypeptide chain. In addition, bacterial BDH isolation was achieved in a two-step purification procedure, improving the knowledge of an enzyme involved in the lipid metabolism of a unique hibernating mammal. Sequence alignment revealed conserved putative amino acids for possible NAD+ interaction.

year	journal	country	edition	language
2008-09-01

https://hal.archives-ouvertes.fr/hal-00374888/file/BMCBiochembdh08.pdf.pdf