Functional analysis of the antigenic structure of a minor T cell determinant from pigeon cytochrome C. Evidence against an alpha-helical conformation.
A minor T cell determinant from pigeon cytochrome c, composed of residues 43 to 58 (p43-58), was synthesized along with a series of 48 analogs containing amino or carboxyl-terminal deletions or single amino acid substitutions. These peptides were analyzed functionally for their ability to elicit unique T cell populations on immunization of C57BL/10 mice and to stimulate a degenerate T cell clone capable of recognizing p43-58 in association with two different Ia molecules, A beta b:A alpha b and A beta d:A alpha d. These experiments allowed us to identify the residues in the determinant that are critical for T cell activation. Residues 50 and 52 had the dominant influence on T cell specificity, and residues 47, 48, 49, 51, and 53 had weak effects. Residues 46 and 54 were hardly recognized by the TCR at all, but appeared to influence the potency of the determinant by interacting with the Ia molecule. Finally, substitutions at positions 55 to 58 had no effect, but removal of these residues reduced the potency of the peptide, suggesting a contribution from the peptide backbone of this part of the molecule during T cell activation. An analysis of the spatial relationship of these dominant epitopic and agretopic residues suggests that this determinant does not assume a pure alpha-helical secondary structure when bound to the Ia molecule.
Ogasawara
K
K
Laboratory of Cellular and Molecular Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Maloy
W L
WL
Beverly
B
B
Schwartz
R H
RH
D000595:Amino Acid Sequence; D000818:Animals; D010856:Columbidae; D003574:Cytochrome c Group / Q000008:administration & dosage / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000008:administration & dosage / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010455:Peptides / Q000008:administration & dosage / Q000276:immunology; D011487:Protein Conformation; D011948:Receptors, Antigen, T-Cell / Q000008:administration & dosage / Q000276:immunology; D013329:Structure-Activity Relationship
D003574:Cytochrome c Group; D000939:Epitopes; D010455:Peptides; D011948:Receptors, Antigen, T-Cell