The fusion of myoblasts to myotubes requires an endogenous soluble metalloendoprotease. To determine whether this protease is released by fusing myoblasts, or stays within the cell, we examined the effects of membrane-impermeant and a membrane-permeant metalloendoprotease inhibitors. Membrane-permeant 1,10-phenanthroline, and membrane-impermeant bathophenanthroline disulfonic acid both inhibited soluble metalloendoprotease activity in homogenized myoblasts with equal potency. However, while 1,10-phenanthroline inhibited fusion, bathophenanthroline disulfonic acid had no effect. In addition, metalloendoprotease activity could not be detected in the media of fusing myoblasts, but was in the cells. These observations support the conclusion that the soluble metalloendoprotease required in fusion remains within the myoblast.