Modulation of peptide binding by HLA-B27 polymorphism in pockets A and B, and peptide specificity of B*2703.

The results in this study address three aspects of peptide binding to the disease-associated antigen HLA-B27 and its modulation by polymorphism: the contribution of major anchor residues 2 and 9, the role of pocket B polymorphism in modulating peptide specificity, and the binding properties of B2703, a subtype not found to be associated with spondyloarthropathy. Synthetic analogs of peptides naturally presented by B2705 were used to demonstrate that residue 2 is essential, since Ala2 analogs bound marginally to B2705, but the specificity of B2705 for Arg2 is not absolute, and show that the contribution of basic residue 9 to binding was significant, but less than Arg2. The effect of single mutations in the B pocket was to decrease or--with the Glu > Met-45 mutation--totally shift pocket B specificity for Arg2 towards other residues at this position. This was shown by quantitating the relative binding of Gln2 and Ala2 analogs, and by pool-sequencing of the peptides bound in vivo to these mutants. Peptides naturally presented by B2705 apparently bound with a lower affinity to pocket A variants with altered hydrogen bonding to the peptide N terminus, including B2703. Binding of peptide analogs with changes at positions 2 or 9 suggested that in B2703 pocket A, interactions are weaker and pocket B interactions are stronger than in B2705. This can be explained by the effect of the unique His59 change in B2703 in both pockets. Thus, B2703 is probably the HLA-B27 sub-type with the most stringent specificity for the Arg2 peptide motif.