Probing HLA-B7 Conformational Shifts Induced by Peptide-Binding Groove Mutations and Bound Peptide with Anti-HLA Monoclonal Antibodies

Kelly D. Smith, Brian E. Mace, Alicia Valenzuela, Janet L. Vigna, Jane McCutcheon, James A. Barbosa, Eric Huczko, Victor H. Engelhard, Charles T. Lutz

Research output: Contribution to journalArticle

Abstract

To determine the influence of peptide-binding groove residues and MHC-bound peptide on HLA-B7 conformation, we investigated the binding sites of nine locus- or allele-specific mAbs using a panel of 82 HLA-B7 variants. The functional mAb epitopes encircle the HLA-B7 peptide-binding groove. Three mAbs are affected by mutations at solvent-accessible peptide-binding groove mutations. Mutations in peptide-binding groove residues 45, 63, and 150 affect multiple nonoverlapping mAb epitopes, probably by interaction with other MHC residues or bound peptide. However, 18 of 24 peptide-binding groove mutations do not affect mAb binding, indicating that the conformation of solvent-accessible HLA-B7 structures is largely dissociated from changes in the peptide-binding groove. To test whether bound peptides alter HLA-B7 conformation, we loaded HLA-B7 heavy chains on acid-stripped cells with β2-microglobulin and 20 individual synthetic peptides. Two of eight mAbs are sensitive to HLA-B7-bound peptides. A likely interpretation of these data is that the conformational flexibility of HLA-B7 is due to peptide-induced conformational shifts in MHC side chains, rather than major shifts in the MHC main chain. These results suggest that HLA-B7 conformation is largely maintained in the context of different bound peptides and different peptide-binding grooves.

Original languageEnglish (US)
Pages (from-to)2470-2478
Number of pages9
JournalJournal of Immunology
Volume157
Issue number6
StatePublished - Sep 15 1996

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HLA-B7 Antigen
Monoclonal Antibodies
Peptides
Mutation
Epitopes

ASJC Scopus subject areas

  • Immunology

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Smith, K. D., Mace, B. E., Valenzuela, A., Vigna, J. L., McCutcheon, J., Barbosa, J. A., ... Lutz, C. T. (1996). Probing HLA-B7 Conformational Shifts Induced by Peptide-Binding Groove Mutations and Bound Peptide with Anti-HLA Monoclonal Antibodies. Journal of Immunology, 157(6), 2470-2478.

Probing HLA-B7 Conformational Shifts Induced by Peptide-Binding Groove Mutations and Bound Peptide with Anti-HLA Monoclonal Antibodies. / Smith, Kelly D.; Mace, Brian E.; Valenzuela, Alicia; Vigna, Janet L.; McCutcheon, Jane; Barbosa, James A.; Huczko, Eric; Engelhard, Victor H.; Lutz, Charles T.

In: Journal of Immunology, Vol. 157, No. 6, 15.09.1996, p. 2470-2478.

Research output: Contribution to journalArticle

Smith, KD, Mace, BE, Valenzuela, A, Vigna, JL, McCutcheon, J, Barbosa, JA, Huczko, E, Engelhard, VH & Lutz, CT 1996, 'Probing HLA-B7 Conformational Shifts Induced by Peptide-Binding Groove Mutations and Bound Peptide with Anti-HLA Monoclonal Antibodies', Journal of Immunology, vol. 157, no. 6, pp. 2470-2478.
Smith, Kelly D. ; Mace, Brian E. ; Valenzuela, Alicia ; Vigna, Janet L. ; McCutcheon, Jane ; Barbosa, James A. ; Huczko, Eric ; Engelhard, Victor H. ; Lutz, Charles T. / Probing HLA-B7 Conformational Shifts Induced by Peptide-Binding Groove Mutations and Bound Peptide with Anti-HLA Monoclonal Antibodies. In: Journal of Immunology. 1996 ; Vol. 157, No. 6. pp. 2470-2478.
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