Hydrolase activities in normoblasts of beta-thalassemic patients

A physiological role for glycosidases in cell membranes has been suggested. Therefore the activities of four glycosidases--beta-galactosidase, beta-glucoronidase, N-acetyl-beta-glucosaminidase and acid phosphatase--were examined in normoblasts and membranes of red blood cells (RBC). The enzymatic assays were based on the hydrolysis of fluorimetric 4-methylumbelliferone from the enzyme substrate. In order to avoid contamination by lysosomal activities derived from RBC, the mature RBC and normoblasts obtained from normal controls and thalassemic patients were separated from other blood elements by cellulose chromatography. The cells were disrupted and lysed by freezing and thawing hypotonic solution. Higher enzymatic activities were found in preparations from thalassemic patients than from normal subjects. With a sucrose density gradient, further separation of normoblasts from RBC membranes was obtained, indicating that the normoblast fraction contributed most of the high specific activity found in the thalassemic preparation. It was concluded that relatively high glycosidase activities are present in normoblasts of thalassemic patients. Lower but significant activities were detected in RBC membranes of normal control subjects and thalassemic patients.