Fig. 4.

Binding of Get4/5 to Get3. (A) Tagged wild-type Get4/5 and Get3 are incubated for 2 h at room temperature then bound to Ni-beads. “D” represents incubation in the presence of 2 mM ADP-Mg²⁺, and “T” in the presence of 2 mM ATP-Mg²⁺. (B) His-tagged Get4/5 and Get3 mutants are incubated in the presence of 2 mM ADP-Mg²⁺ as in (A). The YYE/AAA Get4 mutant lacks a TEV cleavage site and is slightly smaller than the other tagged mutants. (C) As in (B), using his-tagged Get3. (D) View from the N-terminal face of Get4. Mutated residues are displayed as spheres with carbons and labels colored based on wild-type level (cyan), weak (yellow), or weakest (red) interactions with Get3. (E) The closed state of Get3 (PDB ID code 2WOJ) with mutated residues displayed and colored as in (D).