TY - JOUR
T1 - The Q-cycle reviewed
T2 - How well does a monomeric mechanism of the bc1 complex account for the function of a dimeric complex?
AU - Crofts, Antony R.
AU - Holland, J. Todd
AU - Victoria, Doreen
AU - Kolling, Derrick R.J.
AU - Dikanov, Sergei A.
AU - Gilbreth, Ryan
AU - Lhee, Sangmoon
AU - Kuras, Richard
AU - Kuras, Mariana Guergova
N1 - Funding Information:
We gratefully acknowledge support for the work reported here from grants NIH RO1 GM35438 (to ARC) and NIH RO1 GM62954 (to SAD).
PY - 2008/7
Y1 - 2008/7
N2 - Recent progress in understanding the Q-cycle mechanism of the bc1 complex is reviewed. The data strongly support a mechanism in which the Qo-site operates through a reaction in which the first electron transfer from ubiquinol to the oxidized iron-sulfur protein is the rate-determining step for the overall process. The reaction involves a proton-coupled electron transfer down a hydrogen bond between the ubiquinol and a histidine ligand of the [2Fe-2S] cluster, in which the unfavorable protonic configuration contributes a substantial part of the activation barrier. The reaction is endergonic, and the products are an unstable ubisemiquinone at the Qo-site, and the reduced iron-sulfur protein, the extrinsic mobile domain of which is now free to dissociate and move away from the site to deliver an electron to cyt c1 and liberate the H+. When oxidation of the semiquinone is prevented, it participates in bypass reactions, including superoxide generation if O2 is available. When the b-heme chain is available as an acceptor, the semiquinone is oxidized in a process in which the proton is passed to the glutamate of the conserved -PEWY- sequence, and the semiquinone anion passes its electron to heme bL to form the product ubiquinone. The rate is rapid compared to the limiting reaction, and would require movement of the semiquinone closer to heme bL to enhance the rate constant. The acceptor reactions at the Qi-site are still controversial, but likely involve a "two-electron gate" in which a stable semiquinone stores an electron. Possible mechanisms to explain the cyt b150 phenomenon are discussed, and the information from pulsed-EPR studies about the structure of the intermediate state is reviewed. The mechanism discussed is applicable to a monomeric bc1 complex. We discuss evidence in the literature that has been interpreted as shown that the dimeric structure participates in a more complicated mechanism involving electron transfer across the dimer interface. We show from myxothiazol titrations and mutational analysis of Tyr-199, which is at the interface between monomers, that no such inter-monomer electron transfer is detected at the level of the bL hemes. We show from analysis of strains with mutations at Asn-221 that there are coulombic interactions between the b-hemes in a monomer. The data can also be interpreted as showing similar coulombic interaction across the dimer interface, and we discuss mechanistic implications.
AB - Recent progress in understanding the Q-cycle mechanism of the bc1 complex is reviewed. The data strongly support a mechanism in which the Qo-site operates through a reaction in which the first electron transfer from ubiquinol to the oxidized iron-sulfur protein is the rate-determining step for the overall process. The reaction involves a proton-coupled electron transfer down a hydrogen bond between the ubiquinol and a histidine ligand of the [2Fe-2S] cluster, in which the unfavorable protonic configuration contributes a substantial part of the activation barrier. The reaction is endergonic, and the products are an unstable ubisemiquinone at the Qo-site, and the reduced iron-sulfur protein, the extrinsic mobile domain of which is now free to dissociate and move away from the site to deliver an electron to cyt c1 and liberate the H+. When oxidation of the semiquinone is prevented, it participates in bypass reactions, including superoxide generation if O2 is available. When the b-heme chain is available as an acceptor, the semiquinone is oxidized in a process in which the proton is passed to the glutamate of the conserved -PEWY- sequence, and the semiquinone anion passes its electron to heme bL to form the product ubiquinone. The rate is rapid compared to the limiting reaction, and would require movement of the semiquinone closer to heme bL to enhance the rate constant. The acceptor reactions at the Qi-site are still controversial, but likely involve a "two-electron gate" in which a stable semiquinone stores an electron. Possible mechanisms to explain the cyt b150 phenomenon are discussed, and the information from pulsed-EPR studies about the structure of the intermediate state is reviewed. The mechanism discussed is applicable to a monomeric bc1 complex. We discuss evidence in the literature that has been interpreted as shown that the dimeric structure participates in a more complicated mechanism involving electron transfer across the dimer interface. We show from myxothiazol titrations and mutational analysis of Tyr-199, which is at the interface between monomers, that no such inter-monomer electron transfer is detected at the level of the bL hemes. We show from analysis of strains with mutations at Asn-221 that there are coulombic interactions between the b-hemes in a monomer. The data can also be interpreted as showing similar coulombic interaction across the dimer interface, and we discuss mechanistic implications.
KW - Constraints on molecular mechanism
KW - Coulombic interaction
KW - Kinetic model
KW - Q-cycle
KW - Thermodynamic model
KW - bc complex
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U2 - 10.1016/j.bbabio.2008.04.037
DO - 10.1016/j.bbabio.2008.04.037
M3 - Review article
C2 - 18501698
AN - SCOPUS:46449118774
SN - 0005-2728
VL - 1777
SP - 1001
EP - 1019
JO - Biochimica et Biophysica Acta - Bioenergetics
JF - Biochimica et Biophysica Acta - Bioenergetics
IS - 7-8
ER -