Polymorphism at position 97 in MHC class I molecules affects peptide specificity, cell surface stability, and affinity for beta2-microglobulin.
The two mouse MHC class I alleles, L(d) and L(q), share complete amino acid sequence identity except in the alpha2 domain, where they differ at six positions. Despite their similarity, L(q) has a stronger association with beta2-microglobulin (beta2m), is expressed at higher levels on the cell surface, demonstrates an increased cell surface half-life, and has fewer open forms on the cell surface than L(d). To determine the basis for their phenotypic differences, L(d) molecules containing chimeric L(d)-L(q) alpha2 domains were characterized, and these analyses implicated residue 97 (L(d)Trp and L(q)Arg) as the polymorphic site responsible for the disparity in beta2m association between the two alleles. Single substitution analysis at this site (L(d)W97R and L(q)R97W) confirmed this. Furthermore, the L(d)W97R mutant molecule has a longer cell surface half-life than either L(q) or L(d), and fewer open forms of L(d)W97R are observed on the cell surface. In addition, both L(d)W97R and L(q) possess decreased binding affinity for the L(d)-restricted tum(-) P91A(14-22) peptide compared with L(d). Collectively, these results and the known location of Trp(97) in the peptide binding cleft of L(d) strongly suggest that the substitution of Arg for Trp(97) in L(d) alters the peptide binding cleft, increasing its affinity for endogenous peptides, which results in greater cell surface stability and better retention of beta2m. Furthermore, these results imply that Trp(97) plays an important role in the ability of L(d) to efficiently participate in alternative MHC class I Ag presentation pathways.
Smith
Ruth A
RA
Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, MO 65212, USA.
Myers
Nancy B
NB
Robinson
Melanie
M
Hansen
Ted H
TH
Lee
David R
DR
D000483:Alleles; D019943:Amino Acid Substitution / Q000235:genetics / Q000276:immunology*; D000818:Animals; D001120:Arginine / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000939:Epitopes / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D006183:H-2 Antigens / Q000235:genetics* / Q000378:metabolism; D006207:Half-Life; D064532:Histocompatibility Antigen H-2D; D007519:Isoantigens / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010455:Peptides / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D011110:Polymorphism, Genetic / Q000276:immunology*; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D014162:Transfection; D014364:Tryptophan / Q000235:genetics; D001613:beta 2-Microglobulin / Q000378:metabolism*
D000939:Epitopes; D006183:H-2 Antigens; D064532:Histocompatibility Antigen H-2D; D007519:Isoantigens; D010455:Peptides; D001613:beta 2-Microglobulin; D014364:Tryptophan; D001120:Arginine