Molecular and Cellular Biochemistry

, Volume 77, Issue 2, pp 103–110 | Cite as

Hemin-mediated oxidative inactivation of 3-hydroxy-3-methylglutaryl CoA reductase

  • S. Usha Devi
  • T. Ramasarma
Original Article

Summary

Addition of hemoglobin, methemoglobin, hemin or hematin in the assay mixture of rat liver 3-hydroxy-3-methylglutaryl CoA (HMGCoA) reductase inhibited the activity of the enzyme. The inhibition by hemin was rapid, without any apparent dependence on time of preincubation. At 20 μM hemin, a maximum of about 50% inhibition was obtained in the case of the microsomal enzyme while the solubilized enzyme showed almost 80%6 inhibition. Dithiothreitol at high concentrations or either of the two substrates of the enzyme (HMGCoA and NADPH) could afford partial protection when added before hemin. The Km for both the substrates increased in the presence of hemin. The inhibition by hemin appeared to be irreversible, the presence of KCN or NaN3 being the only means of preventing the inhibition. Molecular oxygen was required for the inhibition. Oxygen radicals and H2O2, however, did not seem to be involved. This offered a clue that an oxidation reaction of the reductase protein may be the likely mechanism of its inactivation. The enzyme protein did not, however, get degraded under the conditions of inhibition.

Key words

HMGCoA reductase hemin oxidative inactivation 

Abbreviations

HMGCoA

3-Hydroxy-3-methylglutaryl coenzyme

DTT

Dithiothreitol

DTNB

5,5′-Dithiobis-(2-nitrobenzoic acid)

SDS-PAGE

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis

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Copyright information

© Martinus Nijhoff Publishers 1987

Authors and Affiliations

  • S. Usha Devi
    • 1
  • T. Ramasarma
    • 1
  1. 1.Department of BiochemistryIndian Institute of ScienceBangaloreIndia

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