Cleavage efficiency at limiting substrate concentrations.

<p>The concentration of indicated protease fragments and the substrates xLC3B-MBP (left) or xGATE16-MBP (right) was titrated at constant protease/substrate ratio (1:1000 or 1:2000, respectively). After cleavage (1 h at 0°C), a fraction of each reaction corresponding to 1.2 μg (≈20 pmol) of substrate protein was analyzed by SDS-PAGE. Due to the different substrate concentrations, the absolute volume of the cleavage reaction analyzed by SDS-PAGE had to be adjusted accordingly.