Rapid Electron Transfer within the III-IV Supercomplex in Corynebacterium glutamicum.
Complex III in C. glutamicum has an unusual di-heme cyt. c1and it co-purifies with complex IV in a supercomplex. Here, we investigated the kinetics of electron transfer within this supercomplex and in the cyt. aa3alone (cyt. bc1was removed genetically). In the reaction of the reduced cyt. aa3with O2, we identified the same sequence of events as with other A-type oxidases. However, even though this reaction is associated with proton uptake, no pH dependence was observed in the kinetics. For the cyt. bc1-cyt. aa3supercomplex, we observed that electrons from the c-hemes were transferred to CuAwith time constants 0.1-1 ms. The b-hemes were oxidized with a time constant of 6.5 ms, indicating that this electron transfer is rate-limiting for the overall quinol oxidation/O2reduction activity (~210 e-/s). Furthermore, electron transfer from externally added cyt. c to cyt. aa3was significantly faster upon removal of cyt. bc1from the supercomplex, suggesting that one of the c-hemes occupies a position near CuA. In conclusion, isolation of the III-IV-supercomplex allowed us to investigate the kinetics of electron transfer from the b-hemes, via the di-heme cyt. c1and heme a to the heme a3-CuBcatalytic site of cyt. aa3.
Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.|Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
Simone
S
Graf
Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
Olga
O
Fedotovskaya
Institute of Biochemistry and Molecular Biology, ZBMZ, Faculty of Medicine, BIOSS Centre for Biological Signalling Studies, University of Freiburg, 79104 Freiburg, Germany.
Wei-Chun
WC
Kao
Institute of Biochemistry and Molecular Biology, ZBMZ, Faculty of Medicine, BIOSS Centre for Biological Signalling Studies, University of Freiburg, 79104 Freiburg, Germany.
Carola
C
Hunte
Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
Pia
P
Ädelroth
IBG-1: Biotechnology, Institute of Bio- and Geosciences, Forschungszentrum Jülich, Wilhelm-Johnen-Strasse, D-52425 Jülich, Germany.
Michael
M
Bott
Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
Christoph
C
von Ballmoos
Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
Peter
P
Brzezinski
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