Polarity of the P1 anchor residue determines peptide binding specificity between HLA-A*3101 and HLA-A*3303.

A previous pool sequence analysis showed that HLA-A3101 and HLA-A3303 binding peptides have the same anchor residues at P2 and the C-terminus, the only difference being that HLA-A3303 binding peptides have two additional P2 anchor residues. Using a stabilization assay with RMA-S transfectants expressing HLA-A3101 and human beta2-microglobulin, we tested the binding of 232 8- to 11-mer peptides carrying HLA-A3303 anchor residues to HLA-A3101. One hundred of these peptides (43.1%) bound to HLA-A3101, confirming that these residues are also anchors for HLA-A3101. Although aromatic hydrophobic P2 residues were previously shown to be stronger anchors than aliphatic hydrophobic P2 residues in HLA-A3303 binding peptides, we detected no significant difference in HLA-A3101 binding affinity between peptides carrying aromatic or aliphatic hydrophobic P2 residues. Statistical analysis previously showed a positive effect of negatively charged P1 residues and a negative effect of positively charged P1 residues for peptide binding to HLA-A3303. In contrast such analysis demonstrated a positive effect of positively charged P1 residues and a negative effect of negatively charged P1 residues for peptide binding to HLA-A3101. Analysis using mutated peptides confirmed these results. The present study therefore demonstrates that peptide binding specificity between HLA-A3101 and HLA-A3303 is determined by the polarity of the P1 anchor residue.