NMR-derived model for a peptide-antibody complex

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Abstract
The TE34 monoclonal antibody against cholera toxin peptide 3 (CTP3, VEVPGSQHIDSQKKA) was sequenced and investigated by two-dimensional transferred NOE difference spectroscopy and molecular modeling. The VH sequence of TE34, which does not bind cholera toxin, shares remarkable homology to that of TE32 and TE33, which are both anti-CTP3 antibodies that bind the toxin. However, due to a shortened heavy chain CDR3, TE34 assumes a radically different combining site structure. The assignment of the combining site interactions to specific peptide residues was completed by use of AcIDSQRKA, a truncated peptide analogue in which lysine-13 was substituted by arginine, specific deuteration of individual polypeptide chains of the antibody, and a computer model for the Fv fragment of TE34. NMR-derived distance restraints were than applied to the calculated model of the Fv to generate a three-dimensional structure off the TE34/CTP3 complex. The combining site was found to be a very hydrophobic cavity composed of seven aromatic residues. Charged residues are found in the peripheral of the combining site. The peptide residues HIDSQKKA form a .beta.-turn inside the combining site. The contact area between the peptide and the TE34 antibody is 388 .ANG.2, about half of the contact area observed in protein-antibody complexes.

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