Two-dimensional NMR investigations of the interactions of antibodies with peptide antigens.
To increase our understanding of the molecular basis for antibody specificity and for the cross-reactivity of anti-peptide antibodies with native proteins it is important to study the three-dimensional structure of antibody complexes with their peptide antigens. For this purpose it may not be necessary to solve the structure of the whole antibody complex but rather to concentrate on elucidating the combining site structure, the interactions of the antibody with its antigen and the bound peptide conformation. We have developed an NMR methodology based on two-dimensional difference spectrum measurements which extract the information concerning antibody-peptide interactions and intramolecular interactions in the bound ligand from the crowded and unresolved spectrum of the Fab complex. These measurements yield restraints on interproton distances in the complex which are used to dock the peptide into calculated models for the antibodies' combining sites. Comparison of the interactions of three antibodies against a cholera toxin peptide (CTP3), which differ in their cross-reactivity with the toxin, yields information about the size and conformation of antigenic determinants recognized by antibodies, the structure of their combining sites and relationships between antibodies' primary structure, and their interactions with peptide antigens.
Anglister
J
J
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
Scherf
T
T
Zilber
B
B
Levy
R
R
Zvi
A
A
Hiller
R
R
Feigelson
D
D
D000595:Amino Acid Sequence; D000936:Antigen-Antibody Complex; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D002772:Cholera Toxin / Q000276:immunology*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*; D011487:Protein Conformation
D000936:Antigen-Antibody Complex; D010446:Peptide Fragments; C050310:cholera toxin, B subunit (50-64); D002772:Cholera Toxin