FIGURE 4.
Determination of TRAMs in binding RNA. (A) RNA-EMSA (rEMSA) was performed to assay the sequence specificity of TRAM3066-binding RNA. Twelve of the RNA Pentaprobes (PP) (Bendak et al., 2012) were 3′ biotinylated and used as substrates. Purified TRAM3066 was added to the binding mixture at the indicated concentrations. Arrows point to the RNA probe and its concentration. (B) Surface plasmon resonance (SPR) assayed the binding affinity of TRAM3066 and TRAM2002 to the selected RNA PP1 and PP10. E. coli CspA is included as a reference. Calculated KD values are shown in the corresponding diagrams for each pair of protein-RNAs. The protein concentrations for each reaction are color-coded and labeled in the bottom rows. s, second.