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. 2012 Aug 28;7(8):e41731. doi: 10.1371/journal.pone.0041731

Figure 3. Cross-linking of AG22 constructs with 14-3-3.

Figure 3

A. Cross-linking of truncation constructs AG22 (1–422) S16D/S411D, AG22 (38–422) (S411D) and AG22 (1–145) S16D with 14-3-3. For each cross-linking reaction, the mixture of AG22 protein construct and 14-3-3 or the individual AG22 proteins was incubated with BS3 for 10 min at room temperature. The mixtures before and after cross-linking were analyzed by SDS-PAGE and visualized by Coomassie Blue staining. Lanes on the left show BS3 cross-linking results with 14-3-3; lanes on the right show BS3 cross-linking results for AG22 proteins without 14-3-3. *indicates the cross-linked complex of 14-3-3 with AG22 (1–422) S16D/S411D; §the cross-linked complex of 14-3-3 with AG (38–422) S411D and #the cross-linked complex of 14-3-3 with AG22 (1–145) S16D. B. No detectable cross-linking was observed for AG22 (167–422) S411D and 14-3-3. The arrow indicates the position of the band (90 kDa) expected if AG22 (164–422) S411D (MW, 31.3 kDa) and 14-3-3 (monomer MW, 29.3 kDa) had formed a cross-linked complex. Cross-linking experiments shown in panels A and B are representative of three replicates.