Extensive alanine substitutions increase binding affinity of an influenza nucleoprotein peptide to HLA-Aw68 and do not abrogate peptide-specific CTL recognition.

Abstract

Class I MHC molecules bind peptides in the endoplasmic reticulum and present them at the cell surface to circulating CD8+ T cells for analysis. We have examined binding of peptides and stabilization of HLA-Aw68 class I molecules using synthetic peptide variants of an influenza virus nucleoprotein peptide, NP91-99 (KTGGPIYKR). We have demonstrated that insertion of increasing numbers of alanines in the center of the peptide (between P and I), to increase a natural bulging out of the peptide-binding cleft, results in a large decrease in thermal stability. Although there is a great decrease in the t 1/2 of the MHC/peptide complex for NP-1A compared with NP91-99, a T cell line, stimulated by NP91-99, recognizes NP-1A efficiently. Peptide variants with three or more alanines do not show saturable binding to HLA-Aw68 and also are not recognized by the T cell line. Thermal studies show that polyalanine peptides with minimal anchors and nearly all TCR contact residues exchanged stabilized HLA-Aw68 to high temperatures. Additionally, some of these polyalanine peptides are recognized by T cell lines generated against NP91-99. Analysis of the peptide sequences show that the stabilization effects are not due to the hydrophobicity of the peptide. These data suggest that the strength of binding of peptides to HLA-Aw68 is not only dictated by the presence of anchor residues but also by the lack of unfavorable contacts between the peptide ligand and class I MHC-binding cleft.

• Authors: E J Collins, B L Booth Jr, V Cerundolo
• Journal: Journal of immunology (Baltimore, Md. : 1950) (J Immunol) Vol. 162 Issue 1 Pg. 331-7 (Jan 01 1999) ISSN: 0022-1767 [Print] United States
• PMID: 9886403 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Biopolymers
• Epitopes, T-Lymphocyte
• HLA-A Antigens
• HLA-A*68 antigen
• Nucleoproteins
• Peptide Fragments
• Peptides
• Viral Core Proteins
• polyalanine
• Alanine

Topics

• Alanine (metabolism)
• Amino Acid Sequence
• Amino Acid Substitution (immunology)
• Biopolymers (biosynthesis, immunology)
• Cytotoxicity Tests, Immunologic
• Epitopes, T-Lymphocyte (chemistry, immunology, metabolism)
• HLA-A Antigens (biosynthesis, chemistry, metabolism)
• Humans
• Influenza A virus (immunology)
• Molecular Sequence Data
• Nucleoproteins (immunology, metabolism)
• Peptide Fragments (chemical synthesis, immunology, metabolism)
• Peptides (metabolism)
• Protein Binding (immunology)
• T-Lymphocytes, Cytotoxic (immunology, metabolism)
• Viral Core Proteins (immunology, metabolism)