SPECIES-SPECIFIC DIFFERENCES IN CHAPERONE INTERACTION OF HUMAN AND MOUSE MAJOR HISTOCOMPATIBILITY COMPLEX CLASS-I MOLECULES

Previous studies have shown that immature mouse class I molecules transiently associate with a resident endoplasmic reticulum protein of 88 kD that has been proposed to act as a chaperone for class I assembly. Subsequently this protein was demonstrated to be identical to calnexin and to associate with immature forms of the T cell receptor complex, immunoglobulin, and human class I HLA heavy chains. In this paper we define further the interaction of human class I HLA heavy chains with chaperone proteins and find key differencess with the complexes observed in the mouse system. First, calnexin and immunoglobulin binding protein (BiP) both associate with immature HLA class I heavy chains. The two chaperones are not found within the same molecular complex, suggesting that calnexin and BiP do not interact simultaneously with the same HLA class I heavy chain. Second, only free HLA class I heavy chains, and not beta(2)-microglobulin (beta(2)m)-associated heavy chains are found associated with the chaperones. Indeed, addition of free beta(2)m in vitro induces dissociation of chaperone-class I HLA heavy chain complexes. The kinetics for dissociation of the class I HLA heavy chain-chaperone complexes and for formation of the class I HLA heavy chain-beta(2)m complex display a reciprocity that suggests the interactions with chaperone and beta(2)m are mutually exclusive. Mouse class I heavy chains expressed in human cells exhibit the mouse pattern of interaction with human chaperones and human Pam and not the human pattern, showing the difference in behavior is purely a function of the class I heavy chain sequence.