Further improvement in preparation and some properties of native alpha and beta chains from canine hemoglobin treated with p-chloromercuribenzoate.
The alpha and beta chains were prepared from canine hemoglobin by a modification of the method of Bucci and Fronticelli ((1965) J. Biol. Chem. 240, PC551-552) which involved treatment of hemoglobin with an excess of p-chloromercuribenzoate (pCMB), separation of the mercurated chains by chromatography on a DE-32 column with a salt gradient at pH 8.6, and regeneration of sulfhydryl groups of the chains with 2-mercaptoethanol. The SH titer was two per heme for both the regenerated alpha and beta chains. The titer decreased to four per tetramer of hemoglobin after equimolar recombination of both chains. Measurements of the absorption spectrum and oxygen binding showed that the chains were in a native state.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002729:Chloromercuribenzoates; D004285:Dogs; D004592:Electrophoresis, Starch Gel; D006418:Heme / Q000032:analysis; D006454:Hemoglobins / Q000302:isolation & purification*; D006801:Humans; D046911:Macromolecular Substances; D011487:Protein Conformation; D013045:Species Specificity; D013053:Spectrophotometry; D013438:Sulfhydryl Compounds / Q000032:analysis; D020245:p-Chloromercuribenzoic Acid
D000596:Amino Acids; D002729:Chloromercuribenzoates; D006454:Hemoglobins; D046911:Macromolecular Substances; D013438:Sulfhydryl Compounds; D006418:Heme; D020245:p-Chloromercuribenzoic Acid