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Red cell diphosphoglycerate mutase. Immunochemical studies in vertebrate red cells, including a human variant lacking
Abstract:
Diphosphoglycerate mutase (DPGM) was purified to homogeneity from human erythrocytes. The enzyme and Freund adjuvant were injected 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.
Insights
Diphosphoglycerate mutase (DPGM) is present in neonatal red blood cells across species, but absent in adult goats. A genetic variant lacks DPGM activity and shows altered enzyme stability.
Area of Science:
- Biochemistry
- Immunology
- Hematology
Background:
- Diphosphoglycerate mutase (DPGM) is a crucial enzyme in red blood cells.
- Understanding DPGM distribution and characteristics is important for hematological research.
Purpose of the Study:
- To purify DPGM from human erythrocytes.
- To develop a monospecific antibody for DPGM detection.
- To investigate DPGM presence and characteristics in various species and in a genetic variant.
Main Methods:
- Enzyme purification and antibody production.
- Radial immunodiffusion and Ouchterlony analysis for DPGM quantification and comparison.
- Immunoelectrophoresis and stability assays for characterizing the genetic variant.
Main Results:
- DPGM was purified to homogeneity from human erythrocytes.
- A monospecific antibody against DPGM was successfully generated.
- DPGM was detected in neonatal red cells of dogs, rabbits, rats, chickens, and goats, but not in adult goats.
- Human red cell DPGM levels were consistent between young and old cells.
- A patient with a DPGM genetic variant showed reduced DPGM levels, altered immunoelectrophoretic properties, and decreased enzyme stability.
Conclusions:
- DPGM is present in neonatal erythrocytes of multiple species, with a notable absence in adult goats.
- The developed antibody is effective for quantifying DPGM and identifying variants.
- The DPGM genetic variant exhibits reduced enzyme levels and altered stability, contributing to erythrocytosis.