Purification and properties of l-3-glycerophosphate dehydrogenase from pig brain mitochondria
- 15 October 1980
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 192 (1) , 9-18
- https://doi.org/10.1042/bj1920009
Abstract
L-3-Glycerophosphate dehydrogenase (EC 1.1.99.5) was purified from pig brain mitochondria by extraction with deoxycholate, ion-exchange chromatography and (NH4)2SO4 fractionation in cholate, and preparative isoelectric focusing in Triton X-100. Sodium dodecyl sulfate/polyacrylamide-gel electrophoresis revealed the purified enzyme consisted of a single subunit of MW 75,000. The enzyme contained non-covalently bound FAD and low concentrations of Fe and acid-labile sulfide. No substrate-reducible EPR signals were detected. The conditions of purification, particularly the isoelectric-focusing step, lead to considerable loss of FAD and possibly Fe-S centers, so determining whether the enzyme was a flavoprotein or a ferroflavoprotein was not possible. The enzyme catalyzes the oxidation of L-3-glycerophosphate by a variety of electron acceptors, including ubiquinone analogues. A number of compounds that inhibit ubiquinone oxidoreduction by other enzymes of the respiratory chain failed to inhibit L-3-glycerophosphate dehydrogenase, except at very high concentrations.This publication has 29 references indexed in Scilit:
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