Iron-sulphur centres in mitochondria from Arum maculatum spadix with very high rates of cyanide-resistant respiration

Abstract

X-band electron-paramagnetic-resonance spectroscopy at 4.2–77K combined with measurements of oxidation-reduction potential was used to identify iron–sulphur centres in Arum maculatum (cuckoo-pint) mitochondria. In the oxidized state a signal with a derivative maximum at g = 2.02 was assigned to succinate dehydrogenase centre S-3. Unreduced particles showed additional signals at g = 2.04 and 1.98 (at 9.2 GHz), which may be due to a spin-spin interaction. In the reduced state a prominent signal at g = 1.93 and 2.02 was resolved into at least three components that could be assigned to centres S-1 and S-2 of succinate dehydrogenase (midpoint potentials −7 and −240 mV respectively at pH 7.2) and a small amount of centre N-1b (e'o= −240 mV) of NADH-ubiquinone reductase. In addition, changes in line shape around −10 mV indicated the presence of a fourth component in this signal. The latter was more readily reduced by NADH than by succinate, suggesting that it might be associated with the external NADH dehydrogenase. The iron-sulphur centres of NADH-ubiquinone reductase were present in an unusually low concentration, indicating that the alternative, non-phosphorylating, NADH dehydrogenase containing a low number of iron-sulphur centres may be responsible for most of the high rate of oxidation of NADH.