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class I downregulation represents a significant challenge for successful T cell-based immunotherapy. T cell epitopes associated with impaired peptide processing (TEIPP) constitute a novel category of immunogenic Ags that are selectively presented on transporter associated with Ag processing-deficient cells. The TEIPP neoepitopes are CD8 T cell targets, derived from nonmutated self-proteins that might be exploited to prevent immune escape. In this study, the crystal structure of H-2D(b) in complex with the first identified TEIPP Ag (MCLRMTAVM) derived from the Trh4 protein has been determined to 2.25 \u00c5 resolution. In contrast to prototypic H-2D(b) peptides, Trh4 takes a noncanonical peptide-binding pattern with extensive sulfur-\u03c0 interactions that contribute to the overall complex stability. Importantly, the noncanonical methionine at peptide position 5 acts as a main anchor, altering only the conformation of the H-2D(b) residues Y156 and H155 and thereby forming a unique MHC/peptide conformer that is essential for recognition by TEIPP-specific T cells. Substitution of peptide residues p2C and p5M to the conservative \u03b1-aminobutyric acid and norleucine, respectively, significantly reduced complex stability, without altering peptide conformation or T cell recognition. In contrast, substitution of p5M to a conventional asparagine abolished recognition by the H-2D(b)/Trh4-specific T cell clone LnB5. We anticipate that the H-2D(b)/Trh4 complex represents the first example, to our knowledge, of a broader repertoire of alternative MHC class I binders.","bibjson":{"author":[{"initials":"I","lastname":"Hafstrand","name":"Hafstrand I"},{"initials":"EM","lastname":"Doorduijn","name":"Doorduijn EM"},{"initials":"AD","lastname":"Duru","name":"Duru AD"},{"initials":"J","lastname":"Buratto","name":"Buratto J"},{"initials":"CC","lastname":"Oliveira","name":"Oliveira CC"},{"initials":"T","lastname":"Sandalova","name":"Sandalova T"},{"initials":"T","lastname":"van Hall","name":"van Hall T"},{"initials":"A","lastname":"Achour","name":"Achour A"}],"identifier":[{"id":"10.4049/jimmunol.1502249","type":"doi"},{"id":"26800871","type":"pubmed"}],"issue":[null],"journal":{"iso_abbreviation":"J. 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