Hemoglobin/O2 systems: mechanistic discrimination based on Ackers' model.

Abstract

The kinetics of the reaction of hemoglobin with molecular oxygen, in which rapid mixing is followed by a fast temperature jump, is numerically simulated. We use the system of Ackers (1998) which distinguishes four forms of bi-ligated hemoglobin. The data suggest the involvement of isomerization steps for bi- and triliganded hemoglobin. Our first model assumes a linear addition of oxygen with one path to and from each bi-ligated species. Our second model allows cross-overs between paths, as described by Ackers (1998). Our third model exploits the observation (Perrella et al., 1990) that two of the four bi-ligated forms are at low concentration. We explore whether these models can be distinguished experimentally. We find a narrow oxygen concentration range where Models 1 and 2 can be distinguished by rapid flow experiments. The distinction between Models 2 and 3 is larger in stopped flow experiments within a limited oxygen concentration range but not easily detectable in chemical relaxation following rapid flow. The detection of two special states of free hemoglobin may be possible at low oxygen concentration. However, the step reaction free enthalpy (or Gibbs free energy) values make it more likely that two special states are present in fully ligated hemoglobin.