Changes in acid proteolytic activity during differentiation of murine erythroleukemic cells.
Proteolytic activity was measured in murine erythroleukemic 745 cell line grown in culture, before and after the addition of agents which promote differentiation. The 36,000 X g soluble fraction of the cells degraded [14C]globin with maximal activity at pH 3.6, while the insoluble fraction failed to degrade [14C]globin within a pH range of 2.5-9.0. The acid protease activity in the soluble fraction of the undifferentiated murine erythroleukemic cells increased during the first 2 days in culture and remained constant during the following 4 days. We suggest that this activity resides in the lysosomes since it migrates together with the lysosomal marker alpha-mannosidase on colloidal silica gradients, shows maximum activity at acid pH and is sensitive towards inhibition by pepstatin. Induced differentiation of the cells by dimethyl sulfoxide, butyric acid or hexamethylene bisacetamide was concomitantly associated with a marked reduction in protease activity and the accumulation of hemoglobin within the cells. In contrast, in a non-inducible variant of 745 cell line DMSO failed to affect proteolysis. It is suggested that in murine erythroleukemic cells changes in acid protease activity are associated with the cellular triggered by chemical inducers.
D000818:Animals; D002250:Carbon Radioisotopes; D002454:Cell Differentiation; D002460:Cell Line; D005914:Globins / Q000378:metabolism; D007700:Kinetics; D007942:Leukemia, Experimental / Q000503:physiopathology*; D051379:Mice; D010447:Peptide Hydrolases / Q000378:metabolism*
D002250:Carbon Radioisotopes; D005914:Globins; D010447:Peptide Hydrolases