Single amino acid changes in DR and antigen define residues critical for peptide-MHC binding and T cell recognition.
Single amino acid substitutions of Ag and MHC were used to analyze the fine structure of the influenza hemagglutinin (HA)-derived epitope (HA 307-319) recognized in the context of DR7 molecules by a T cell clone. Putative T cell (HA 308, 310, 311, 313, and 316) and DR (HA 309, 312, and 317) contact residues of the Ag were identified by the use of single amino acid-substituted analogs that were tested for their T cell-activating and DR-binding capacities. The peptide-DR7-T cell interaction was further characterized by the use of a panel of 13 site-directed DR7 mutant transfectants analyzed for their capacity to present Ag to T cells, and for their purified mutant DR7 molecules to bind HA 307-319 or its single amino acid-substituted analogs. Eight mutants lost their Ag-presenting function, whereas only one had any decrease in peptide binding. Finally, for three of the mutants it was possible to correct the deleterious effects of mutation by using a particular single amino acid-substituted analog of the peptide molecule. The observed pattern of complementation led to a model that predicts that the Ag assumes an extended conformation, with a turn, in the binding groove, such that the following residues are in close proximity: DR 86-HA 309, DR 71-HA 312, DR 30-HA 314, and 315.
Krieger
J I
JI
Cytel, La Jolla, CA 92037.
Karr
R W
RW
Grey
H M
HM
Yu
W Y
WY
O'Sullivan
D
D
Batovsky
L
L
Zheng
Z L
ZL
Colón
S M
SM
Gaeta
F C
FC
Sidney
J
J
D000595:Amino Acid Sequence; D000938:Antigen-Presenting Cells / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D001665:Binding Sites; D002999:Clone Cells; D000939:Epitopes; D006684:HLA-DR Antigens / Q000737:chemistry / Q000378:metabolism*; D006389:Hemagglutinins, Viral / Q000276:immunology*; D006801:Humans; D007111:Immunity, Cellular; D066298:In Vitro Techniques; D008285:Major Histocompatibility Complex; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000276:immunology*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000276:immunology*
D000956:Antigens, Viral; D000939:Epitopes; D006684:HLA-DR Antigens; D006389:Hemagglutinins, Viral