PMID: 6411123Aug 16, 1983Paper

Effect of auto-oxidized phospholipids on oxidative enzyme assays based on tetrazolium salt reduction

Biochimica Et Biophysica Acta
E M Massa, R N Farías

Abstract

The influence of auto-oxidized phospholipids on the reduction of the tetrazolium salt MTT coupled to the NAD+-dependent lactate dehydrogenase reaction was studied. The following results were obtained: (1) peroxidized phosphatidylcholine interfered in the time-course of the lactate dehydrogenase-mediated MTT reduction; (2) there was a time-dependent decrease in the hydroperoxide content of phosphatidylcholine vesicles during the incubation; (3) the diminution of phosphatidylcholine hydroperoxides required the presence of all the components of the system except MTT; (4) hydroperoxide diminution and MTT reduction were mediated by the superoxide radical O2-, since both processes were inhibited by superoxide dismutase; (5) EDTA inhibited the hydroperoxide decrease and abolished the interference of peroxidized phosphatidylcholine with MTT reduction. It was concluded that hydroperoxides compete with MTT for the electrons coming from substrate oxidation. The superoxide radical O2- and traces of some contaminating metal ion are involved in the process. This is a potential complication in the study of the effect of lipids on enzymatic activities assayed by the tetrazolium salt method.

References

Jan 1, 1978·Methods in Enzymology·J A Buege, S D Aust
Jan 31, 1972·Biochemical and Biophysical Research Communications·M NishikimiK Yagi
Apr 16, 1973·Biochimica Et Biophysica Acta·S Batzri, E D Korn
Nov 1, 1971·Analytical Biochemistry·C Beauchamp, I Fridovich
Sep 16, 1981·Biochemical and Biophysical Research Communications·G W RuddockC L Greenstock
Mar 1, 1982·Archives of Biochemistry and Biophysics·M W Sutherland, J M Gebicki

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Citations

Oct 1, 1988·The Histochemical Journal·C J Van Noorden
Feb 14, 2002·Biochemical Pharmacology·Avikam HarelKonstantin Bloch
Jun 3, 2016·Microbiology and Molecular Biology Reviews : MMBR·David R Soll, Karla J Daniels

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