Figure 2. SDS-PAGE analysis of VHH inhibition of GST-SNAP cleavage by BoNT/A Lc.
(A) Inhibitory effect of 15 unique VHH fragments. Each VHH was incubated in a 50 fold molar excess over BoNT/A Lc for 3 min followed by the addition of GST-SNAP25141-206. After 10 min of incubation, SNAP25 cleavage was analyzed by SDS-PAGE. (B) Effect of molar ratio on GST-SNAP25 cleavage. Two inhibitory and one non-inhibitory VHH were incubated with varying fold molar excesses over BoNT/A Lc (1:1 to 70:1) for 30 min followed by the addition of GST-SNAP25141-206. After 10 min of incubation, SNAP25 cleavage was analyzed by SDS-PAGE.