The isolation and partial characterization of diphosphoglycerate mutase from human erythrocytes.
Diphosphoglycerate mutase has been purified to homogeneity from outdated human erythrocytes. The native enzyme has a molecular weight of 57 000 as determined by equilibrium centrifugation and exclusion chromatography. Disc gel electrophoresis in the presence of sodium dodecyl sulfate yields a single protein band with a molecular weight of about 26 500, indicating that diphosphoglycerate mutase is comprised of two subunits of similar mass. The enzyme exhibits the following intrinsic activities: diphosphoglyceratemutase, monophosphoglycerate mutase, and 2,3-diphosphoglycerate phosphatase. The latter activity is enhanced in the presence of either organic or inorganic anions. Glycolate-2-P, particularly, has a profound activating effect. Nonspecific phosphatase and enolase activities are absent. The enzyme has an extinction coefficient at 280 nm of 1.65 cm2/mg. The amino acid composition of the homogeneous protein has been determined.
D000596:Amino Acids / Q000032:analysis; D001731:Bisphosphoglycerate Mutase / Q000097:blood* / Q000302:isolation & purification; D004912:Erythrocytes / Q000201:enzymology*; D006461:Hemolysis; D006358:Hot Temperature; D006801:Humans; D008970:Molecular Weight; D010770:Phosphotransferases / Q000097:blood*
D000596:Amino Acids; D010770:Phosphotransferases; D001731:Bisphosphoglycerate Mutase