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. 2019 May 3;10:966. doi: 10.3389/fimmu.2019.00966

Figure 5.

Figure 5

Experimental validation of peptide-modulation of HLA-A2 α3 domain motions via steady state fluorescence anisotropy. (A) The HLA-A2 protein and the fluorescent label in the 220s loop of the α3 domain (loop in green; residues 220–226). The binding of the CD8 coreceptor is illustrated to show its relationship to the α3 domain and the 220s loop (44). (B) Fluorescence anisotropy (reported in millianisotropy values) measured for D220C-labeled HLA-A2 bound to five different nonameric peptides. For calibration, a fully rigid molecule has a theoretical value of 400, and free fluorescein had a value of < 10. Measurements are the averages and standard deviations from analysis of three independently prepared samples. A single asterisk indicates differences between the Tax sample and the WT1 and Flu M1 samples with p < 0.05. The double asterisk indicates a difference between the Tax sample and the gp1002M with p < 0.0005. (C) Comparison of the measurements for the five peptide/HLA-A2 samples in panel A with the RMS fluctuations at position 220 from the molecular dynamics simulations.