Identification of residues necessary for clonally specific recognition of a cytotoxic T cell determinant.
The residues in an influenza nucleoprotein (NP) cytotoxic T cell determinant necessary for cytotoxic T cell (CTL) recognition, were identified by assaying the ability of hybrid peptides to sensitize a target cell to lysis. The hybrid peptides were formed by substituting amino acids from one determinant (influenza NP 147-158) for the corresponding residues of a second peptide (HLA CW3 171-182) capable of binding to a common class I protein (H-2Kd). Six amino acids resulted in partial recognition; however, the presence of a seventh improved the potency of the peptide. Five of the six amino acids were shown to be required for recognition. The spacing of the six amino acids was consistent with the peptide adopting a helical conformation when bound. The importance of each amino acid in CTL recognition and binding to the restriction element was investigated further by assaying the ability of peptides containing point substitutions either to sensitize target cells or to compete with the natural NP sequence for recognition by CTL. The T cell response was much more sensitive to substitution than the ability of the peptide to bind the restriction element. Collectively the separate strategies identified an approximate conformation and orientation of the peptide when part of the complex and permitted a potential location in the MHC binding site to be identified. The model provides a rationalization for analogues which have previously been shown to exhibit greater affinity for the class I molecule and suggests that the binding site in major histocompatibility complex (MHC) class I molecules might have greater steric constraints that the corresponding area of class II proteins.
Rothbard
J B
JB
Laboratory of Molecular Immunology, Imperial Cancer Research Fund, London, UK.
Pemberton
R M
RM
Bodmer
H C
HC
Askonas
B A
BA
Taylor
W R
WR
D000595:Amino Acid Sequence; D000818:Animals; D000939:Epitopes; D005805:Genes, MHC Class I; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D009154:Mutation; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D011487:Protein Conformation; D011948:Receptors, Antigen, T-Cell; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014758:Viral Core Proteins / Q000276:immunology
D000939:Epitopes; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D011948:Receptors, Antigen, T-Cell; D014758:Viral Core Proteins