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Biophys J, June 2001, p. 2622-2630, Vol. 80, No. 6

Temperature Transition of Human Hemoglobin at Body Temperature: Effects of Calcium

Christina Kelemen,* Shu Chien,dagger and G. M. Artmann*

 *Department of Cell Biophysics, University of Applied Sciences Aachen, D-52428 Juelich, Germany; and  dagger Whitaker Institute of Biomedical Engineering and Department of Bioengineering, University of California, San Diego, La Jolla, California 92093 USA

We studied the effects of calcium ion concentration on the temperature dependence of rheological behavior of human red blood cells (RBCs) and concentrated hemoglobin solutions. Our previous study (G. M. Artmann, C. Kelemen, D. Porst, G. Büldt, and S. Chien, 1998, Biophys. J., 75:3179-3183) showed a critical temperature (Tc) of 36.4 ± 0.3°C at which the RBCs underwent a transition from non-passage to passage through 1.3-µm micropipettes in response to an aspiration pressure of -2.3 kPa. An increase in intracellular Ca2+ concentration by using the ionophore A23187 reduced the passability of intact RBCs through small micropipettes above Tc; the micropipette diameter needed for >90% passage increased to 1.7 µm. Viscometry of concentrated hemoglobin solutions (45 and 50 g/dl) showed a sudden viscosity transition at 36 ± 1°C (Tceta ) at all calcium concentrations investigated. Below Tceta , the viscosity value of the concentrated hemoglobin solution at 1.8 mM Ca2+ was higher than that at other concentrations (0.2 µM, 9 mM, and 18 mM). Above Tceta , the viscosity was almost Ca2+ independent. At 1.8 mM Ca2+ and 36 ± 1°C, the activation energy calculated from the viscometry data showed a strong dependence on the hemoglobin concentration. We propose that the transition of rheological behavior is attributable to a high-to-low viscosity transition mediated by a partial release of the hemoglobin-bound water.

Biophys J, June 2001, p. 2622-2630, Vol. 80, No. 6
© 2001 by the Biophysical Society   0006-3495/01/06/2622/09  $2.00


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