The charge transfer events in the SA(L223) reaction center mutant Rhodobacter sphaeroides chromatophores were investigated by direct electrometry. Besides the primary charge separation, the small stigmatellin-sensitive electrogenic reaction due to the electron transfer from the primary to the secondary quinone acceptor in the reaction center complex was observed after the first flash. The second flash-induced electrogenic phase of the secondary quinone protonation and subsequent electrogenic reactions of the cytochrome bc 1 complex were much slower than those in chromatophores of the wild type. It is suggested that replacement of Ser-L223 by Ala impairs both specific proton-conducting pathways leading to the secondary quinone Q B .