alpha-chymotrypsin and benzidine

The addition of certain proteases to cultures of Friend virus-infected mouse erythroleukemia cells can induce up to 90% of the cells in culture to become hemoglobin-containing, as assessed by positive staining for benzidine (B+). Because the mechanism of this protease action is unknown, media components were studied as possible targets for protease activity. Aliquots of medium plus serum were incubated for various times with levels of protease sufficient to induce approximately 50% of the cells to the B+ state. Cells were added to protease-pretreated serum either before or after inactivation of the protease. In all cases, enzymatically active protease had to be present with the cells to induce B+ cells to form. Serum and other components of the medium pretreated with protease were inactive. Mouse erythroleukemia cells grown in the absence of serum were also induced by proteases to form B+ cells. These data imply that the inducing action of proteases cannot be passively transferred by protease-pretreated serum or medium nor is serum required for protease-mediated induction of B+ cells. Taken together, these conclusions suggest that the protease action is on the cells or on cellular products intimately associated with cells.

Topics: Animals; Benzidines; Blood; Cell Differentiation; Cell Line; Chymotrypsin; Culture Media; Dimethyl Sulfoxide; Friend murine leukemia virus; Hemoglobins; Leukemia, Erythroblastic, Acute; Mice; Peptide Hydrolases