Thermodynamic volume cycles for electron transfer in the cytochrome c oxidase and for the binding of cytochrome c to cytochrome c oxidase.
Dilatometry is a sensitive technique for measuring volume changes occurring during a chemical reaction. We applied it to the reduction-oxidation cycle of cytochrome c oxidase, and to the binding of cytochrome c to the oxidase. We measured the volume changes that occur during the interconversion of oxidase intermediates. The numerical values of these volume changes have allowed the construction of a thermodynamic cycle that includes many of the redox intermediates. The system volume for each of the intermediates is different. We suggest that these differences arise by two mechanisms that are not mutually exclusive: intermediates in the catalytic cycle could be hydrated to different extents, and/or small voids in the protein could open and close. Based on our experience with osmotic stress, we believe that at least a portion of the volume changes represent the obligatory movement of solvent into and out of the oxidase during the combined electron and proton transfer process. The volume changes associated with the binding of cytochrome c to cytochrome c oxidase have been studied as a function of the redox state of the two proteins. The volume changes determined by dilatometry are large and negative. The data indicate quite clearly that there are structural alterations in the two proteins that occur on complex formation.
Kornblatt
J A
JA
Enzyme Research Group, Departments of Biology, Chemistry and Biochemistry, Concordia University, Montréal, Québec H3G 1M8, Canada. krnbltt@bax2.concordia.ca
Kornblatt
M J
MJ
Rajotte
I
I
Hoa
G H
GH
Kahn
P C
PC
D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D003574:Cytochrome c Group / Q000737:chemistry* / Q000378:metabolism*; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000378:metabolism*; D066298:In Vitro Techniques; D008956:Models, Chemical; D010084:Oxidation-Reduction; D011485:Protein Binding; D011522:Protons; D013816:Thermodynamics
D003574:Cytochrome c Group; D011522:Protons; D003576:Electron Transport Complex IV