Polymerization of hemoglobin in sickle trait erythrocytes and lysates.

C.T Noguchi,D.A Torchia,A.N Schechter

doi:10.1016/s0021-9258(19)69413-5

Abstract

We report 13C/1H magnetic double resonance measurements of polymer in sickle trait erythrocytes and direct measurements of solubility on lysates of these cells as a function of oxygen saturation. Polymerized hemoglobin is detected by these two methods only at oxygen saturation values below 60% and increases to 0.4 of the total hemoglobin at complete deoxygenation. These results confirm the validity of the NMR approach in determining values of polymer fraction. We have also found that the thermodynamic theory, that successfully describes gelation in sickle erythrocytes, does not predict the experimental results for sickle trait erythrocytes and must be refined.

Highlights

AS erythrocytes were prepared from blood obtained from individuals with sickletrait
We have found that the thermodynamic theory, that successfully describes gelation in sickle erythrocytes, does not predict the experimental results for sickle trait erythrocytes and must be refined
Intracellular gelation in AS erythrocytes should be nearly identical with gelationof comparable solutions of hemoglobin A and S mixtures

Summary

Polymerization of Hemoglobin in Sickle Trait Erythrocytes and Lysates*

We have measured the amount of polymer formed within AS erythrocytes as a function of oxygen saturation using 13C/’H double resonance methods [11]. T h e solubility of AS cell lysate was (Received for publication, December 12, 1980, and in revised form, February 20, 1981). From the §Laboratory of Chemical Biology, National Institute ofrlrthritis, Metabolism, and Digestive Diseases and the VLaboratory of Biochemistry, National lnstitute measured as a function of oxygen saturation using ultracentrifuge sedimentation. The amountsof polymer predicted for these samplesusing the thermodynamic theoryof Minton [7] were calculated and compared with the experimental measurements, as were data on thesickling behavior of AS erythrocytes

MATERIALS AND METHODS

Polymerization in Sickle Trait Erythrocytes

DISCUSSION