The function of the subunits of the fumarate reductase complex of Vibrio succinogenes

Abstract

The membrane-bound fumarate reductase complex of Vibrio succinogenes catalyzes the reduction of fumarate by 2,3-dimethyl-1,4-naphthohydroquinone (dimethylnaphthohydroquinone) and consists of three different peptides (Mr 79,000, Mr 31,000 and Mr 25,000), the smallest of which is cytochrome b [Unden, G., Hackenberg, H. and Kröger A. (1980) Biochem. Biophys. Acta 591, 275-288]. The complex was cleaved with guanidinium chloride, the resulting subunits characterized and their functions within the complex investigated by reconstitutional experiments. 1. The Mr-79,000 subunits catalyzed the reduction of fumarate by benzylviologen radicals as well as the oxidation of succinate by methylene blue, but not fumarate reduction by dimethylnaphthohydroquinone. 2. The spectral and the redox properties of the isolated cytochrome b (Mr 25,000) were equivalent to those of the high-potential cytochrome b of the bacteria. The isolated cytochrome b had a midpoint potential of -15 mV and was reducible by dimethylnaphthohydroquinone in the absence of the other subunits. 3. The Mr-31,000 subunit did not catalyze any of the reactions mentioned above. For the reduction of cytochrome b by succinate in the presence of the Mr-79,000 subunit, an amount of the Mr-31,000 subunit was required which was equimolar to cytochrome b. 4. The activity of fumarate reduction by dimethylnaphthohydroquinone could be restored by coprecipitation of the three subunits. It is concluded that the fumarate reductase complex has two different sites, which are essential for its function in the phosphorylative electron transport of the bacterium. The site reacting with the substrates fumarate and succinate is situated on the Mr-79,000 subunit, and that reacting with dimethylnaphthohydroquinone is cytochrome b. The Mr-31,000 subunit mediates the electron transport between cytochrome b and the Mr-79,000 subunit.