Assay of [3H] Dihydrorotenone Binding to Complex I in Intact Human Platelets

F. Blandini, J. T. Greenamyre

Research output: Contribution to journalArticle

Abstract

We have developed an assay for the binding of [3H]dihydrorotenone ([3H]DHR), an analogue of the pesticide rotenone, to the mitochondrial enzyme, complex I, in intact human platelets. The highly hydrophobic nature of dihydrorotenone, which diffuses easily through biological membranes, rendered the isolation of mitochondrial fractions unnecessary. This allowed us to reduce the amount of blood required and to shorten the processing of samples considerably. [3H]DHR binding was saturable, specific, and highly reproducible. We also found that MPP+ (1-methyl-4-phenylpyridinium species), which is accumulated actively by platelets, inhibited [3H]DHR specific binding in a concentration-dependent manner. This method could provide a simple tool for the study of complex I in those disorders, such as Parkinson′s disease (PD), in which a defect of this enzyme has been suggested.

Original languageEnglish
Pages (from-to)16-19
Number of pages4
JournalAnalytical Biochemistry
Volume230
Issue number1
DOIs
Publication statusPublished - Sep 1995

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Biophysics
  • Biochemistry

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