Fig. 6
Analysis of the temperature-dependent interaction between C13Fv and THETAS. a Flowchart depicting the analysis. Predicted structures of C13Fv and THETAS were used for docking simulation together with amino acid information of THETAS. Among the resultant 248 docked structures, nine structures (d) were selected in which five or six important amino acids (b, c) are involved in the binding. b Affinity constants between C13 mAb and THETAS mutants measured by indirect competitive ELISA at 4, 15, 26, and 37 °C. Error bars represent standard deviation (n = 3). c Epitope analysis by western blot of mutated THETAS. Overall pictures are shown in Supplementary Fig. 2c, d. d Superimposed drawing of nine selected docking structures. H chain, L chain, CDRs of H chain, and CDRs of L chain are colored in light orange, light pink, orange, and purple, respectively. Nine predicted structures of THETAS are differently colored. e, f Temporal changes in averaged RMSDs of THETAS (e) or C13Fv (f) in the MD simulation. g Heatmap showing the RMSDs from 5 ns prior to THETAS dissociation in the MD simulation. Dissociation of THETAS from C13Fv was judged when the average value of RMSD of α carbon for each amino acid of THETAS exceeded 20 Å by a simple expedient
