Peptide-independent stabilization of MHC class I molecules breaches cellular quality control.

ABSTRACT

The 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 α-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 (ß2m), and bypasses all three cellular quality control steps. Our data demonstrate (1) that coupling between peptide and ß2m binding to the MHC-I heavy chain is mediated by conformational dynamics; (2) that the folded conformation of MHC-I, supported by ß2m, plays a decisive role in passing the ER-to-cell-surface transport quality controls; and (3) that ß2m association is also tested by the cell surface quality control that leads to MHC-I endocytosis.