High-pressure laser photolysis study of hemoproteins. Effects of pressure on carbon monoxide binding dynamics for R- and T-state hemoglobins.
The bimolecular association reaction of carbon monoxide to human adult hemoglobin at pH 7, 20 degrees C, was examined as a function of pressure up to 1500 bar by means of high-pressure laser photolysis. The apparent quantum yield for a millisecond recombination reaction decreased with pressure, which was attributed to an increase in the fraction of nanosecond geminate recombination reaction. On the basis of the pressure dependence of the recombination rate, the activation volumes at normal pressure for the binding of carbon monoxide to the R- and T-state hemoglobins were determined as -9.0 +/- 0.7 and -31.7 +/- 2.4 cm3 mol-1, respectively. Since the activation volumes for the overall CO association reaction were negative, it seems that the iron-ligand bond formation process mainly contributes to the rate-limiting step for both quaternary structures. The characteristic pressure dependence of the activation volume was observed for the R-state Hb but not for the T-state Hb. At 1000 bar, the activation volume for the R-state Hb was reduced to nearly zero, probably resulting from the contribution of the ligand migration process to the rate-limiting step. The effect of pressure on the activation enthalpy and entropy was also extracted from the data.
Unno
M
M
Division of Molecular Engineering, Graduate School of Engineering, Kyoto University, Japan.
Ishimori
K
K
Morishima
I
I
D001274:Atmospheric Pressure; D002248:Carbon Monoxide / Q000378:metabolism*; D002263:Carboxyhemoglobin / Q000032:analysis* / Q000528:radiation effects; D006454:Hemoglobins / Q000378:metabolism* / Q000528:radiation effects; D006801:Humans; D007834:Lasers; D010777:Photochemistry; D011485:Protein Binding; D013816:Thermodynamics
D006454:Hemoglobins; D002248:Carbon Monoxide; D002263:Carboxyhemoglobin