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. 2014 Jan 9;426(1):199–214. doi: 10.1016/j.jmb.2013.09.016

Fig. 7.

Fig. 7

Differential binding of biotinylated molecules by divalent streptavidins. (a) A monobiotinylated PCR product (50 nM) was incubated with 1,3 trans- or cis-divalent streptavidin (100 nM) for the indicated times at 37 °C and then analyzed by agarose gel electrophoresis, with ethidium bromide visualization of DNA. C is a control of DNA without streptavidin. (b) Titration of the biotinylated PCR product (50 nM) with varying amounts of 1,3 trans-divalent (upper panel) or cis-divalent (lower panel) streptavidin for 16 h at 25 °C, analyzed by electrophoresis as in (a). (c) Native PAGE of monobiotinylated affibody titrated with 1,3 trans-divalent (upper panel) or cis-divalent (lower panel) streptavidin, analyzed by Coomassie staining. (d) Comparison of the relative sizes of an affibody (PDB ID: 2KZJ, space-fill), a short segment of B-form DNA (PDB ID: 1BNA, space-fill), and the separation between cis-binding sites on streptavidin (PDB ID: 3RY2, protein in gray, biotin in red, Connolly surface).