Photoacoustic characterization of protein dynamics following CO photodetachment from fully reduced bovine cytochrome c oxidase.
We report a protein conformational change following carbon monoxide photodetachment from fully reduced bovine cytochrome c oxidase that is hypothesized to be associated with changes in ligand mobility through a dioxygen access channel in the protein. Although not resolved by earlier photoacoustic or optical studies on this adduct, utilization of slightly lower temperatures revealed a process with a kinetic lifetime of about 70 ns at 10 degrees C. We measure an enthalpy change of about 8 kcal/mol in 0.050 M HEPES buffer that becomes less endothermic (DeltaH approximately 2 kcal/mol) at higher ionic strength. The volume contraction of about -0.7 mL/mol associated with the process almost doubles in higher ionic strength buffer systems. Measurements of samples in phosphate buffer systems are similar and appear to display the same subtle ionic strength dependence. Both the isolation of this photoacoustic signal component and the possible dependence on ionic strength of the thermodynamic parameters derived from its analysis appear analogous to and consistent with prior photoacoustic results monitoring CO photodetachment from the camphor complex of cytochrome P-450. Accordingly, we consider a similar model in which a conformational change results in movement of an exposed charged group or groups towards the interior of the protein, out of contact with solvent, as in the closing of a salt bridge.
Oertling
W Anthony
WA
Department of Chemistry and Biochemistry, 226 Science Building, Eastern Washington University, Cheney, WA 99004-2440, USA. WOertling@ewu.edu
Cornellison
Charisa D
CD
Treff
Nathan R
NR
Watanabe
Junji
J
Pressler
Michelle A
MA
Small
Jeanne R
JR
D000818:Animals; D002248:Carbon Monoxide / Q000737:chemistry*; D002417:Cattle; D003576:Electron Transport Complex IV / Q000737:chemistry*; D010084:Oxidation-Reduction; D011487:Protein Conformation; D013696:Temperature; D013816:Thermodynamics
D002248:Carbon Monoxide; D003576:Electron Transport Complex IV