A High-Throughput Solid-Phase Microplate Protein-Binding Assay to Investigate Interactions between Myofilament Proteins

Figure 3

A set of typical, original protein-protein binding curves demonstrating the altered binding of an exon 8-excluded cardiac troponin T (exon 8-excluded TnT, white square) to the interacting protein troponin I in comparison with wild-type cardiac troponin T (WT-TnT, black circle). The troponin T variants are configured as the immobilized protein and troponin I the soluble protein. The normalized binding curves demonstrate exon 8-excluded TnT bound with higher affinity to troponin I compared to WT-TnT as evident by the lower concentration required for 50% maximal binding. Absolute absorbance of the maximal binding is shown in the inset demonstrating that both exon 8-excluded TnT and WT-TnT exhibited similar coupling strength to troponin I.