Transient kinetic and oxidation-reduction studies of spinach ferredoxin:nitrite oxidoreductase

Biochim Biophys Acta. 1994 May 18;1185(3):336-45. doi: 10.1016/0005-2728(94)90249-6.


The oxidation-reduction midpoint potentials for the two prosthetic groups of the chloroplast-located, ferredoxin-dependent nitrite reductase of spinach leaves have been determined by spectroelectrochemical titrations and cyclic voltammetry. The average of the results obtained by the two techniques are Em = -290 mV for the siroheme group and Em = -365 mV for the [4Fe-4S] cluster. The value obtained for the [4Fe-4S] cluster is substantially more positive than values obtained previously in experiments which utilized electron paramagnetic resonance spectroscopy at cryogenic temperatures to monitor the reduction state of the cluster. Laser flash photolysis experiments have been used to monitor electron transfer from reduced ferredoxin to nitrite reductase and have provided the first evidence for electron transfer between the two prosthetic groups of the enzyme. The effect of ionic strength on the observed kinetics has provided support for the proposal that electrostatic interactions between ferredoxin and nitrite reductase play an important role in the reaction mechanism.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Kinetics
  • Nitrite Reductases / chemistry*
  • Oxidation-Reduction
  • Plant Proteins / chemistry*
  • Vegetables


  • Plant Proteins
  • Nitrite Reductases
  • ferredoxin-nitrite oxidoreductase