Interactions of the mitochondrial membrane rat liver D-3-hydroxybutyrate dehydrogenase with glass beads during adsorption chromatography. Relationships with the activation of the enzyme by phospholipids.
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.
Adami
P
P
Laboratoire de Biochimie et Biologie Moléculaire (UA CNRS 531), Université de Franche Comté, Besançon, France.
Nasser
B
B
Latruffe
N
N
D000818:Animals; D002850:Chromatography, Gel / Q000295:instrumentation / Q000379:methods*; D004789:Enzyme Activation / Q000502:physiology; D005898:Glass; D006884:Hydroxybutyrate Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D007425:Intracellular Membranes / Q000737:chemistry / Q000201:enzymology*; D008863:Microspheres; D008930:Mitochondria, Liver / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure*; D010743:Phospholipids / Q000378:metabolism / Q000502:physiology*; D051381:Rats
D010743:Phospholipids; D006884:Hydroxybutyrate Dehydrogenase