Studies on the mode of action of diphtheria toxin. VI. Site of the action of toxin in living cells.

Using the technique of radioautography, it has been shown that a probable maximum of only 25-50 molecules iodine-125-labeled toxin per cell is bound by human HeLa cells treated with approximately 10(7) molecules of toxin per cell, or just under one saturating dose. Radioautographs of sections from labeled cells locate most if not all of the toxin molecules fixed to the outer cell membrane. Under identical conditions far less label is taken up by mouse L cells. It is probable that the resistance of this species to diphtheria toxin can be accounted for in terms of the failure of mouse cells to bind the toxin protein. The irreversible inhibition of protein synthesis in a living cell culture by a few molecules of toxin located at the cell surface is discussed in relation to the known interaction between toxin, NAD, and transferase II in mammalian cell extracts.