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intracellular trafficking of major histocompatibility complex class I (MHC-I) proteins is directed by three quality control mechanisms that test for their structural integrity, which is correlated to the binding of high-affinity antigenic peptide ligands. To investigate which molecular features of MHC-I these quality control mechanisms detect, we have followed the hypothesis that suboptimally loaded MHC-I molecules are characterized by their conformational mobility in the F-pocket region of the peptide-binding site. We have created a novel variant of an MHC-I protein, K(b)-Y84C, in which two \u03b1-helices in this region are linked by a disulfide bond that mimics the conformational and dynamic effects of bound high-affinity peptide. K(b)-Y84C shows a remarkable increase in the binding affinity to its light chain, beta-2 microglobulin (\u03b22m), and bypasses all three cellular quality control steps. Our data demonstrate (1) that coupling between peptide and \u03b22m binding to the MHC-I heavy chain is mediated by conformational dynamics; (2) that the folded conformation of MHC-I, supported by \u03b22m, plays a decisive role in passing the ER-to-cell-surface transport quality controls; and (3) that \u03b22m association is also tested by the cell surface quality control that leads to MHC-I endocytosis.","bibjson":{"author":[{"initials":"Z","lastname":"Hein","name":"Hein Z"},{"initials":"H","lastname":"Uchtenhagen","name":"Uchtenhagen H"},{"initials":"ET","lastname":"Abualrous","name":"Abualrous ET"},{"initials":"SK","lastname":"Saini","name":"Saini SK"},{"initials":"L","lastname":"Jan\u00dfen","name":"Jan\u00dfen L"},{"initials":"A","lastname":"Van Hateren","name":"Van Hateren A"},{"initials":"C","lastname":"Wiek","name":"Wiek C"},{"initials":"H","lastname":"Hanenberg","name":"Hanenberg H"},{"initials":"F","lastname":"Momburg","name":"Momburg F"},{"initials":"A","lastname":"Achour","name":"Achour A"},{"initials":"T","lastname":"Elliott","name":"Elliott T"},{"initials":"S","lastname":"Springer","name":"Springer S"},{"initials":"D","lastname":"Boulanger","name":"Boulanger D"}],"identifier":[{"id":"10.1242/jcs.145334","type":"doi"},{"id":"24806963","type":"pubmed"}],"issue":[null],"journal":{"iso_abbreviation":"J. 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