Fig 5. GltB, GltA and GltC interact directly.
(A) GltC interacts with GltB and GltA. Purified MalE, MalE-CglC20-172, MalE-GltB20-275 or MalE-GltA22-256 were mixed with an equal amount of GltC25-673-His6, loaded on an amylose matrix, the matrix was washed and then bound proteins were eluted with 10 mM maltose. Samples before loading (L), from the flow through (FT), the washing step (W) and elution (E) were analyzed by immunoblotting using antibodies against GltC and MalE. In the upper row, arrowheads indicate GltC25-673-His6 with its calculated molecular mass and in the lower row, the individual MalE proteins. Molecular mass markers are indicated to the left. (B) GltB interacts with GltA. Purified MalE, MalE-CglC20-172 or MalE-GltA22-256 were mixed with an equal amount of GST-GltB20-275 and analyzed as described in A. Blots are marked as in A. (C) Summary of direct protein interactions. Green lines indicate interaction detected, red lines indicate interactions tested but not detected; grey line indicates interaction not tested.