Figure 7. Degradation of the catalytic domain of mature hTACE and its prodomain-bound form by the endoproteinase LysC.

hTACE's catalytic domain complex or procatalytic domain complex (8 μg each) was incubated with 0.2 μg of LysC in 200 μl of Tris/HCl containing 150 mM NaCl and 10 μM GW9471 to inhibit the enzymatic activity of TACE. At the specified times, 18 μl of each incubation was processed as specified in the Materials and methods section, then resolved by SDS/PAGE (4–20% gradient gels). (A) hTACE catalytic domain degradation by LysC versus time (min). (B) Degradation of hTACE procatalytic domain complex by LysC versus time (min). The initial positions of undigested pro- and catalytic domains are shown next to the molecular-mass standards.