Figure 6. Effect of NEYSO-9V peptide mutations on the binding of 1G4 TCR to HLA-A2/NYESO-9V.

Side-on view of HLA-A2 in complex with the NYESO peptide with the α2-helix omitted for clarity. Peptide residues in contact with 1G4 TCR are coloured red. Individual peptide residues were mutated to alanine and refolded with WT HLA-A2 heavy chains. The effects of the mutation on affinity and kinetics was determined by SPR and expressed relative to WT as in Table S1. To estimate the total contribution of side chains to binding energy (∑∆∆G) the combined contribution of the adjacent residues M4 and W5 was assumed to be >2.66 kcal/mol. ‘n.m.’ indicates not measureable.