Reconstitution of hemoglobin from isolated peptide chains

Abstract The α and β peptide chains of mouse or rabbit hemoglobin were separated by chromatography of globin on carboxymethylcellulose at acid pH. Rabbit and mouse hemoglobins have been reconstituted from hemin and peptide chains which have been isolated from globin. High recoveries of reconstituted hemoglobin can be obtained by mixing the isolated chains and slowly dialyzing to neutral pH before the addition of hemin. The reconstituted hemoglobin reversibly binds oxygen when reduced to the ferrous form, as does natural hemoglobin. It is possible to form hybrid hemoglobin molecules by reconstituting α or β chains from rabbit mixed with the appropriate peptide chain from mouse globin.