6533b852fe1ef96bd12aaab1

RESEARCH PRODUCT

Interactions of the mitochondrial membrane rat liver d-3-hydroxybutyrate dehydrogenase with glass beads during adsorption chromatography

P. AdamiN. LatruffeB. Nasser

subject

chemistry.chemical_classificationChromatographyOrganic ChemistryPhospholipidDehydrogenaseGeneral MedicineMitochondrionBiochemistryDithiothreitolAnalytical ChemistryEnzyme activatorchemistry.chemical_compoundEnzymeBiochemistrychemistryNAD+ kinaseInner mitochondrial membrane

description

D-3-Hydroxybutyrate dehydrogenase (BDH) is an NAD(+)-dependent dehydrogenase of the mitochondrial inner membrane involved in the energetic balance between the liver and peripheral organs in mammals. It allows the conversion of ketone bodies (acetoacetate and D-3-hydroxybutyrate) and it is one of the best documented lipid-requiring enzymes with a dependence on lecithins. After release of proteins from the membrane by phospholipase A2 treatment of salt-treated mitochondria, the rat liver enzyme is absorbed on controlled-pore glass beads. After batch washing, the enzyme, devoid of lipids (apoBDH), is specifically eluted at pH 8.05-8.15 with a 0.1 M Tris-1 M LiBr buffer under reducing conditions (5 mM dithiothreitol). It appears that during BDH absorption, the glass beads mimic the phospholipid surface of biomembranes.

yearjournalcountryeditionlanguage
1991-01-01Journal of Chromatography A
https://doi.org/10.1016/s0021-9673(01)83936-4