Yasushi Hanakawa, Norman M. Schechter, Chenyan Lin, Luis Garza, Hong Li, Takayuki Yamaguchi, Yasuyuki Fudaba, Koji Nishifuji, Motoyuki Sugai, Masayuki Amagai, John R. Stanley
ETA Amu inhibits cleavage of Dsg1 by ETA. Anti–E-tag immunoblot of hDsg1E (open arrowhead) and its carboxy-terminal cleavage product (filled arrowhead). ETA Amu at a concentration of 13 μM incubated with hDsg1E inhibits cleavage by subsequent addition of wild-type ETA, but at 4 μM does not. At 8 μM, about half of the Dsg1 is free to be cleaved. Kd can then be roughly estimated to be 8 μM. ETA Amu in the concentrations shown (all in large excess of that of hDsg1E) was incubated with hDsg1 at 25°C for 60 minutes, then wild-type ETA was added for a 20-minute incubation before SDS-PAGE.