Figure 5

Nebulin wild-type (WT) super repeat domain affinity for mutant tropomyosins. Purified GST-nebulin domains bound to beads were incubated with purified α-tropomyosin and β-tropomyosin, then beads were washed, and bound proteins were run in SDS-PAGE gels and stained with Coomassie Blue. The relative quantities of bound α-tropomyosin and β-tropomyosin were calculated from three independent experiments. Six β–tropomyosin mutants (p.K7del, p.E41K, p.K49del, p.E117K, p.E139del, and p.Q147P) were included in the study. The mean values and standard deviations from three experiments are shown in the bar chart to the left, and gel pictures of representative experiments are shown on the right. (A) Binding affinities of the β–tropomyosin mutants for nebulin super repeat S9. (B) Binding affinities of the β–tropomyosin mutants for nebulin super repeat S14. (C) Binding affinities of the β–tropomyosin mutants for nebulin super repeat S18. (D) Binding affinities of the β–tropomyosin mutants for nebulin super repeat S22. No statistically significant differences in binding affinities were found. Abbreviations: Neb, nebulin; Tm2, β-tropomyosin; Tm3, α-tropomyosin; GST, glutathione-S-transferase.