Synthesis and intracellular distribution of cathepsins E and D in differentiating murine Friend erythroleukemia cells.
The synthesis, accumulation, and cellular distribution of cathepsins E and D during the dimethyl sulfoxide (DMSO)-induced differentiation of Friend erythroleukemia cells were investigated. The cellular levels of cathepsins E and D rapidly increased within 1 day of DMSO induction and then sharply decreased over the next 7 days. Since the cells during 1 day of differentiation were morphologically the same as uninduced cells, the results suggest the importance of these enzymes in more cellular proteolysis for the following committed differentiation. While cathepsin D was present mostly in the sedimentable fraction of cells throughout the differentiation period, the distribution of cathepsin E varied to the stage of differentiation. The ratio of the soluble/sedimentable cathepsin E content was 1.1, 1.4, 0.9, and 0.7 in cells after 0, 1, 4, and 7 days of DMSO treatment, respectively. The maturation of reticulocytes to erythrocytes was accompanied by complete loss of the soluble cathepsin E and of all of the cellular cathepsin D. Immunoblotting analyses revealed that both uninduced and induced cells contained two forms of cathepsin E; a high molecular weight form (82 kDa) which was mainly associated with the sedimentable fraction and a low molecular weight form (74 kDa) which was found largely in the soluble fraction. The distribution of these two forms was not significantly changed throughout the differentiation period, but the 74-kDa protein was completely eliminated with maturation of reticulocytes to erythrocytes. Cathepsin D also appeared in two forms in both uninduced and induced cells; a minor (46 kDa) and a major (42 kDa) form which appear to have a precursor-product relationship.
Sakai
H
H
Department of Pharmacology, Nagasaki University School of Dentistry, Japan.
Kato
Y
Y
Yamamoto
K
K
D000818:Animals; D002402:Cathepsin D / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D020059:Cathepsin E; D002403:Cathepsins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004905:Erythrocyte Aging; D004910:Erythrocyte Membrane / Q000201:enzymology*; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology* / Q000473:pathology; D007942:Leukemia, Experimental / Q000201:enzymology / Q000473:pathology; D051379:Mice; D008811:Mice, Inbred DBA; D012156:Reticulocytes / Q000201:enzymology*; D013347:Subcellular Fractions / Q000201:enzymology; D014407:Tumor Cells, Cultured
D002403:Cathepsins; D020059:Cathepsin E; D002402:Cathepsin D; D004121:Dimethyl Sulfoxide