The experiments were performed according to our published procedure [29]. To 3 μl of the dissociated protein complex described above, 3 μl each of 10 × G7 Reaction Buffer (0.5 M sodium phosphate/pH 7.5) and 10% NP-40 was added, followed by 21 μl of water. The 30-μl reaction mix was incubated at 37°C overnight in the absence or presence of 1 μl of PNGase F. An equal volume (30 μl) of 2 × Laemmli sample buffer was added to the reaction mix. The 60-μl sample was incubated at 37°C for 10 min and loaded onto SDS-PAGE for separation.
