TY - JOUR
T1 - The bc:caa3 supercomplexes from the Gram positive bacterium Bacillus subtilis respiratory chain
T2 - A megacomplex organization?
AU - Sousa, Pedro M.F.
AU - Videira, Marco A.M.
AU - Santos, Filipe A.S.
AU - Hood, Brian L.
AU - Conrads, Thomas P.
AU - Melo, Ana M.P.
N1 - Funding Information:
We are greatly indebted to Dr Miguel Teixeira for continuous support. The authors would like to thank Dr Lars Hederstedt for the generous gift of the B. subtilis mutant strains. Tiago David, Elisabete Lopes and Isabel Palos are acknowledged for excellent technical assistance. PS is recipient of the FCT Grant SFRH/BD/46553/2008.
PY - 2013
Y1 - 2013
N2 - The respiratory chain of some prokaryotes was shown to be organized in supercomplexes. This association has been proposed to improve enzyme stability and the overall efficiency of the oxidative phosphorylation process. Here, we have revisited recent data on the supercomplexes of Bacillus subtilis respiratory chain, by means of 1D and 2D-BN-PAGE, sucrose gradient fractionation of solubilized membranes, and mass spectrometry analysis of BN-PAGE bands detected in gel for succinate and cytochrome c oxidoreductase activities. The cytochrome bc:caa3 oxygen oxidoreductase supercomplex was observed in different stoichiometries, (bc)4:(caa3)2, (bc) 2:(caa3)4 and 2[(bc)2:(caa 3)4], suggesting for the first time the string association model of supercomplexes in a Gram positive bacterium. In addition, the presence of a succinate:quinone oxidoreductase:nitrate reductase supercomplex was confirmed by the co-localized succinate:nitroblue tetrazolium and methylviologen:nitrate oxidoreductase activities detected in gel and corroborated by LC-MS/MS analysis.
AB - The respiratory chain of some prokaryotes was shown to be organized in supercomplexes. This association has been proposed to improve enzyme stability and the overall efficiency of the oxidative phosphorylation process. Here, we have revisited recent data on the supercomplexes of Bacillus subtilis respiratory chain, by means of 1D and 2D-BN-PAGE, sucrose gradient fractionation of solubilized membranes, and mass spectrometry analysis of BN-PAGE bands detected in gel for succinate and cytochrome c oxidoreductase activities. The cytochrome bc:caa3 oxygen oxidoreductase supercomplex was observed in different stoichiometries, (bc)4:(caa3)2, (bc) 2:(caa3)4 and 2[(bc)2:(caa 3)4], suggesting for the first time the string association model of supercomplexes in a Gram positive bacterium. In addition, the presence of a succinate:quinone oxidoreductase:nitrate reductase supercomplex was confirmed by the co-localized succinate:nitroblue tetrazolium and methylviologen:nitrate oxidoreductase activities detected in gel and corroborated by LC-MS/MS analysis.
KW - Bacillus subtilis
KW - Nitrate reductase
KW - Oxidative phosphorylation
KW - Respiratory chain supercomplex
UR - http://www.scopus.com/inward/record.url?scp=84881120335&partnerID=8YFLogxK
U2 - 10.1016/j.abb.2013.07.012
DO - 10.1016/j.abb.2013.07.012
M3 - Article
C2 - 23880299
AN - SCOPUS:84881120335
SN - 0003-9861
VL - 537
SP - 153
EP - 160
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -