Optimization of the C-terminal ZBTB38 ZFs for use in structural studies with methylated DNA.
A, domain organization of human ZBTB38 and construct design for the C-terminal ZF region used in this study (green boxes). Red boxes depict the N-terminal set of five ZFs, and the blue box designates the BTB/POZ protein:protein interaction domain. B, 1H-15N HSQC spectral overlay for free ZBTB38 ZFs 6–10 (black), ZFs 6–9 (red), and ZFs 7–10 (green). C, 1H-15N HSQC spectral overlay for ZBTB38 ZFs 6–10 (black), ZFs 6–9 (red), and ZFs 7–10 (green) each in complex with the mCZ38BS_21-mer. D, comparative binding isotherms for ZBTB38 ZFs 6–10, 6–9, and 7–10 in complex with mCZ38BS_27-mer. Each data point represents the average of triplicate data, with error bars depicting ± S.D.