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 |
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Authors: | P Adami B Nasser N Latruffe |
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Institution: | Laboratoire de Biochimie et Biologie Moléculaire (UA CNRS 531), Université de Franche Comté, Besan?on, France. |
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Abstract: | 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. |
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