Photoaffinity labelling of mitochondrial NADH dehydrogenase with arylazidoamorphigenin, an analogue of rotenone

Biochem J. 1984 Dec 1;224(2):525-34. doi: 10.1042/bj2240525.


A photoaffinity-labelling analogue of the respiratory inhibitor rotenone was synthesized from the naturally occurring rotenoid amorphigenin. The analogue inhibits NADH-ubiquinone oxidoreductase activity at concentrations comparable with those of rotenone. Photolysis of the radiolabelled analogue bound to isolated NADH-ubiquinone oxidoreductase resulted in preferential incorporation of radioactivity into a polypeptide of Mr 33 000, particularly at low concentrations of the inhibitor. Preparations of the enzyme differ in a parallel fashion in the content of this polypeptide, the degree of photolabelling by the analogue and their sensitivity to rotenone, providing further evidence that the 33 000-Mr protein forms part of the rotenone-binding site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels* / chemical synthesis
  • Animals
  • Azides* / chemical synthesis
  • Cattle
  • Cytochrome Reductases / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Light
  • Macromolecular Substances
  • Mitochondria, Heart / enzymology*
  • NADH Dehydrogenase / antagonists & inhibitors
  • NADH Dehydrogenase / metabolism*
  • Rotenone / analogs & derivatives*
  • Rotenone / chemical synthesis
  • Spectrophotometry, Ultraviolet


  • Affinity Labels
  • Azides
  • Macromolecular Substances
  • Rotenone
  • Cytochrome Reductases
  • NADH Dehydrogenase