The Pib2 complex binds glutamine.

Recombinant 6His-Pib2 or 6His-MBP protein on Ni-NTA agarose was incubated without or with cell lysates of Δpib2 cells (YKOL4391) for 60 min at 4°C. After washing, the [³H]l-glutamine-binding assay was performed as described in Materials and Methods. Unlabeled glutamine was added where indicated. Statistical data are shown as Mean ± SE of three independent experiments. ****p < 0.0001, Student’s t-test. The purified proteins were also separated by SDS-PAGE and visualized by Coomassie staining (right panel).