Purification of bisphosphoglyceromutase, 2,3-bisphosphoglycerate phosphatase and phosphoglyceromutase from human erthrocytes. Three enzyme activities in one protein.
Bisphosphoglyceromutase, 2,3-bisphosphoglycerate phosphatase and phosphoglyceromutase have been purified from human red cells. Three enzymes were co-purified throughout all purification steps. Three fractions (peaks I, II and III) which were chromatographically separable and had three activities in different ratios were obtained. Peak III which contained the main bisphosphoglyceromutase and 2,3-bisphosphoglycerate phosphatase activities was purified to homogeneity by electrophoretic and ultracentrifugal analyses. The homogeneous preparation had the phosphoglyceromutase activity. The three activities were lost at the same rate during thermal inactivation. Thus, bisphosphoglyceromutase and 2,3-bisphosphoglycerate phosphatase activities, which are responsible for 2,3-bisphosphoglycerate metabolism in red cells, are displayed by the same enzyme protein which has phosphoglyceromutase activity. Peaks I and II were rich in the phosphoglyceromutase activity. Both peaks showed bisphosphoglyceromutase and 2,3-bisphosphoglycerate phosphatase activities, although these two activities were much smaller than those of peak III. Some of the enzymic properties of peak III are described. Comparative studies on three peaks showed that the phosphoglyceromutase of peak III differed from that of peaks I and II in the kinetic property and thermostability.
D002845:Chromatography; D002850:Chromatography, Gel; D004163:Diphosphoglyceric Acids; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D006882:Hydroxyapatites; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000097:blood / Q000302:isolation & purification*; D010770:Phosphotransferases / Q000097:blood / Q000302:isolation & purification*; D013816:Thermodynamics; D014461:Ultracentrifugation
D004163:Diphosphoglyceric Acids; D006882:Hydroxyapatites; D010770:Phosphotransferases; D010744:Phosphoric Monoester Hydrolases