The stability of the heme-globin linkage in some normal, mutant, and chemically modified hemoglobins.
The rates and equilibria of heme exchange between methemoglobin and serum albumin were measured using a simple new spectrophotometric method. It is based on the large difference between the spectrum of methemoglobin and that of methemealbumin at pH 8-9. The rate of heme exchange was found to be independent of the albumin concentration and inversely proportional to the hemoglobin (Hb) concentration. Taken together with the finding that the rate was 10 times greater for Hb Rothschild, which is completely dissociated into alpha beta dimers and 10 times smaller for two cross-linked hemoglobins, the subunits of which cannot dissociate, this showed that the rate of dissociation of heme from alpha beta dimers is very much greater than from tetramers. Conditions were found for the attainment of an equilibrium distribution of hemes between beta globin and albumin. The equilibrium distribution ratio, R = methemealbumin/albumin/methemoglobin/apohemoglobin, for hemoglobin A was 3.4 with human and 0.005 with bovine serum albumin. Both the rates of exchange and the R values of HbS and HbF were the same as that for HbA. The equilibrium distribution ratio for Hb Rothschild was 7 times greater than that for HbA whereas that of one but not the other of the cross-linked hemoglobins was 10 times smaller. The structural bases for these differences are analyzed.
Benesch
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Department of Biochemistry and Molecular Biophysics, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Kwong
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D005260:Female; D005312:Fetal Blood; D005319:Fetal Hemoglobin / Q000378:metabolism; D005914:Globins / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D006441:Hemoglobin A / Q000302:isolation & purification / Q000378:metabolism; D006451:Hemoglobin, Sickle / Q000302:isolation & purification / Q000378:metabolism; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism*; D006455:Hemoglobins, Abnormal / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D009154:Mutation; D011247:Pregnancy; D011485:Protein Binding
D006451:Hemoglobin, Sickle; D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; D006418:Heme; D005914:Globins; D006441:Hemoglobin A; D005319:Fetal Hemoglobin