Comparison of the state of deoxyhemoglobin S molecules in solution and in fibers by hydrogen exchange kinetics

Abstract

Hydrogen exchange kinetics of deoxyhemoglobin S gel and deoxyhemoglobin A solution were compared at 4.8 m m tetramer concentration, 25 °C, and in sodium phosphate buffer at pH 7.0 with T 2 = 0.2 by means of microdialysis using tritium as a trace label. Cyanomethemoglobin A in solution and as crosslinked crystals were compared under the same conditions. The exchange values from 15 to 10 4 min were fitted to a power law, and the distribution function of exchange rates was calculated. There was no significant difference in the distribution for deoxyhemoglobin S gel and deoxyhemoglobin A. Exchange from crosslinked cyanomethemoglobin crystals was less in the early time region than for the solution state, but after 600 min the exchange curves were the same. This resulted in a larger area for the distribution function, although the predominate rates were nearly the same. The effect of polymerization on conformational fluctuations was very small, smaller than the effect of crosslinking hemoglobin crystals.