Figure 3.
Interaction of the UreD–UreF–UreG complex with urease. The following proteins were transiently expressed in N. benthamiana: lane 1, none (control); lane 2, ureD, ureF, and Strep-tagged ureG (DFGS); lane 3, urease (U). The UreD–UreF–UreGS complex was affinity-purified without eluting the proteins from the matrix, and an aliquot was analyzed (lane 4). The clarified extract from urease-expressing leaves was incubated either with purified UreD–UreF–UreGS complex still attached to Streptactin beads or with fresh Streptactin beads (control). After washing, the respective affinity-bound proteins were eluted and detected (lanes 5 and 6) by Western blot analysis. The experiment was repeated two times, and one of the repeats is shown.