Electron transfer kinetics of soluble fragments indicate a direct interaction between complex III and the caa3 oxidase in Thermus thermophilus.
The extremely thermophilic bacterium Thermus thermophilus expresses an aerobic respiratory chain resembling that of mitochondria and many mesophilic prokaryotes. Yet, interaction modes between redox partners differ between the thermophilic and mesophilic electron transport chains. While electron transfer in mesophilic organisms such as Paracoccus denitrificans follows a two-step mechanism mostly governed by long-range electrostatic interactions, the electron transfer in thermophiles is mediated mainly by apolar interactions. The terminal branch of the electron path from the bc-complex via the soluble cytochrome c(552) to the ba(3) oxidase has extensively been characterized, whereas contradicting evidence has been put forward on the nature of the physiological substrate(s) of the caa(3) oxidase. We have cloned and expressed a soluble fragment of the hydrophilic cytochrome c domain derived from subunit IIc of the caa(3) oxidase (c(caa)(3)) and characterized its kinetic behaviour in terms of substrate specificity and ionic strength dependency using pre-steady state stopped-flow techniques. The kinetics revealed fast electron transfer between the caa(3) fragment and both, the cytochrome c(552) and the soluble cytochrome c(bc) fragment of the bc-complex, showing only a weak ionic strength dependence. These data suggest a direct intercomplex electron transfer between the bc-complex and the caa(3) oxidase without requirement for a soluble electron shuttle.
Janzon
Julia
J
Molekulare Genetik, Institut für Biochemie, Biozentrum der J. W. Goethe-Universität, Centre of Excellence Frankfurt Macromolecular Complexes, Frankfurt/Main, Germany.
Ludwig
Bernd
B
Malatesta
Francesco
F
D001483:Base Sequence; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000378:metabolism*; D045264:Cytochromes a / Q000378:metabolism*; D045266:Cytochromes a3 / Q000378:metabolism*; D017931:DNA Primers; D004579:Electron Transport; D014450:Electron Transport Complex III / Q000378:metabolism*; D007700:Kinetics; D012995:Solubility; D016963:Thermus thermophilus / Q000201:enzymology*
D003574:Cytochrome c Group; D045266:Cytochromes a3; D017931:DNA Primers; C435384:cytochrome caa(3); D045264:Cytochromes a; D014450:Electron Transport Complex III