Red Cell

Diphosphoglycerate mutase (DPGM) was purified to homogeneity from human erythrocytes. The enzyme and Freund adjuvant were infected into chickens and yielded a monospecific precipitating antibody. Radial immunodiffusion with this antibody was used to measure the amount of DPGM in hemolysates from human adult and cord red cells. Dog, rabbit, rat, chicken, and goat red cells all had DPGM during the neonatal period, but goat adult red cells had no detectable enzyme. Single bands with no spurs were present on Ouchterlony plates in which human hemolysate was placed adjacent to hemolysates from the other species tested. The amount of human red cell DPGM did not differ between young and old cells separated by centrifugation. Red cells from a patient with a DPGM genetic variant who had erythrocytosis and no detectable enzyme activity contained a reduced amount of DPGM as determined by radial immunodiffusion. The abnormal DPGM differed from normal by immunoelectrophoresis and in stability as measured by the amount of crossreacting material in young versus old erythrocytes. C HANUTIN AND CURNISH’ and Benesch et al.2 showed the importance of 2,3-diphosphoglycerate (DPG), the most abundant organic acid-soluble phosphate in human erythrocytes,3’4 in the control of oxygen binding to hemoglobin. Since then, investigators have isolated the enzyme that catalyzes 2,3DPG production, diphosphoglycerate mutase (DPGM). Sasaki et al.5 isolated homogeneous human erythrocyte DPGM with the aid of isoelectric focusing. Kappel and Hass6 by a less complex technique obtained relatively large amounts of homogeneous human erythrocyte DPGM. A slight modification of the method of Kappel and Hass was used to isolate human DPGM that served as antigen to obtain DPGM-specific antibody from chickens in our laboratory. This paper describes the procedure used to produce DPGM-specific antibody and the results obtained by Ouchterlony and radial immunodiffusion and by imm unoelectrophoresis. Changes during maturation in several vertebrate species were determined. Preliminary results in a patient with no detectable red cell 2,3-DPG and no DPGM activity are also presented.