The Cys-67 residue of HLA-B27 influences cell surface stability, peptide specificity, and T-cell antigen presentation.
Cys-67 of HLA-B27 is located in the B pocket, which determines peptide-binding specificity. We analyzed effects of the Cys-67 --> Ser mutation on cell surface expression, peptide specificity, and T-cell recognition of HLA-B*2705. Surface expression was assessed with antibodies recognizing either native or unfolded HLA proteins. Whereas native B*2705 molecules predominated over unfolded ones, this ratio was reversed in the mutant, suggesting lower stability. Comparison of B*2705- and Cys-67 --> Ser-bound peptides revealed that the mutant failed to bind approximately 15% of the B*2705 ligands, while binding as many novel ones. Two peptides with Gln-2 found in both B*2705 and Cys-67 --> Ser are the first demonstration of natural B*2705 ligands lacking Arg-2. Other effects of the mutation on peptide specificity were: 1) average molecular mass of natural ligands higher than for B*2705, 2) bias against small residues at peptide position (P) 1, and 3) increased P2 permissiveness. The results suggest that the Cys-67 --> Ser mutation weakens B pocket interactions, leading to decreased stability of the mutant-peptide complexes. This may be partially compensated by interactions involving bulky P1 residues. The effect of the mutation on allorecognition was consistent with that on peptide specificity. Our results may aid understanding of the pathogenetic role of HLA-B27 in spondyloarthropathy.
Alvarez
I
I
Centro de Biologia Molecular Severo Ochoa (Consejo Superior de Investigaciones Cientificas), Universidad Autónoma de Madrid, Facultad de Ciencias, 28049 Madrid, Spain.
Martí
M
M
Vázquez
J
J
Camafeita
E
E
Ogueta
S
S
López de Castro
J A
JA
D019943:Amino Acid Substitution; D000818:Animals; D017951:Antigen Presentation; D002460:Cell Line; D000939:Epitopes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D015796:HLA-B27 Antigen / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D009154:Mutation; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
D000939:Epitopes; D015796:HLA-B27 Antigen; D008024:Ligands; D010455:Peptides