The alkylation of hemoglobin S by nitrogen mustard. High resolution proton nuclear magnetic resonance studies.
Sickle cell hemoglobin (Hb S) treated with nitrogen mustard (bis(beta-chloroethyl)methylamine hydrochloride) gives two reaction products, one labile and one stable. After dialysis against buffer solution, the remaining stable product is found to inhibit the polymerization of deoxyhemoglobin S. High resolution proton nuclear magnetic resonance has been used to study the structure and function of this stable product and to investigate the nature of the binding sites of nitrogen mustard to the hemoglobin molecule. The NMR results suggest that the nitrogen mustard treatment of Hb S does not alter the heme environment or the subunit interfaces of the hemoglobin molecule. Moreover, the NMR spectra have also shown that the nitrogen mustard reacts with the beta2 histidines of the hemoglobin molecule and have suggested that several other surface amino acid residues of the hemoglobin molecule are also affected by the nitrogen mustard alkylation. These NMR findings are in good agreement with the data obtained from biochemical studies of nitrogen mustard-treated Hb S. The NMR spectra also indicate that nornitrogen mustard (which is also effective in inhibiting sickling) binds with the hemoglobin molecule in a manner identical with nitrogen mustard. Sulfur mustard, on the other hand, produces no observable changes in the aromatic proton resonances, which is consistent with the fact that it does not inhibit the polymerization of deoxy-Hb S.
D000328:Adult; D001665:Binding Sites; D005021:Ethylamines / Q000097:blood; D006451:Hemoglobin, Sickle; D006455:Hemoglobins, Abnormal; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009588:Nitrogen Mustard Compounds / Q000097:blood; D010100:Oxygen / Q000097:blood; D011485:Protein Binding; D011487:Protein Conformation
D005021:Ethylamines; D006451:Hemoglobin, Sickle; D006455:Hemoglobins, Abnormal; D009588:Nitrogen Mustard Compounds; D010100:Oxygen