Class II-Associated Invariant Chain Peptide-Independent Binding of Invariant Chain to Class II MHC molecules

Abstract

The class II-associated invariant chain peptide (CLIP) region of invariant chain (Ii) is believed to play a critical role in the assembly and transport of MHC class II αβIi complexes through its interaction with the class II peptide-binding site. The role of the CLIP sequence was investigated by using mutant Ii molecules with altered affinity for the DR1 peptide-binding site. Both high- and low-affinity mutants were observed to efficiently assemble with DR1 and mediate transport to endosomal compartments in COS cell transfectants. Using N- and C-terminal truncations, a region adjacent to CLIP within Ii(103–118) was identified that can complement loss of affinity for the peptide-binding site in mediating efficient assembly of αβIi. A C-terminal fragment completely lacking the CLIP region, Ii(103–216), was observed binding stably to class II molecules in immunoprecipitation studies and experiments with purified proteins. The Ii(103–118) region was required for this binding, which occurs through interactions outside of the αβ peptide-binding groove. We conclude that strong interactions involving Ii(103–118) and other regions of Ii cooperate in the assembly of functional αβIi under conditions where CLIP has little or no affinity for the class II peptide-binding site. Our results support the hypothesis that the CLIP sequence has evolved to avoid high-stability interactions with the peptide-binding sites of MHC class II molecules rather than as a promiscuous binder with moderate affinity for all class II molecules.