FIG. 5.

Stachyose reduces the biotinylation of non-surface-located protein MBP, and the biotinylation of KatY is similarly reduced. F1⁻ Y. pestis CO99-3015.S2 and F1⁺ Y. pestis CO99-3015.S1 proteins in the presence or absence of stachyose were biotinylated (or not biotin treated) as in the experiments described in the legends to Fig. 3 and 4. NA-conjugated beads were used to capture biotinylated proteins from the total sonicated cell lysate (for MBP and F1) or from SDS-dissociated total membrane proteins (for KatY and Pla). The bead-bound proteins were boiled in SDS-PAGE sample buffer and analyzed in immunoblots probed with protein-specific antibodies detected by ECL. Flowthrough, proteins that did not bind to the beads; elution, proteins that bound to the beads. The presence (+) versus the absence (−) of stachyose and biotin are indicated.