Data from ¹H nuclear magnetic resonance studies on the Fv fragment of protein 315, a Dnp-binding BALB/c mouse lgA(λ<sub>2</sub>) myeloma protein, have been used to refine a predicted structure of the combining site of the protein. The Dnp-binding subsite in the modified structure is composed of the side chains of three aromatic amino acids Trp 93<sub>L</sub>, Tyr 3<sup>4</sup><sub>L</sub> and 3<sup>4</sup><sub>H</sub>. A fourth aromatic amino acid residue is close to the side chain-NH-CH 2 -group, this is Tyr 33 H - The antibody-hapten binding is a simple encounter process, which causes no extensive conformation change in the Fv fragment. A method for paramagnetic structural studies has been devised using Dnp derivatives with cllgophosphate side chains, which create a specific manganese binding site on the Fv fragment-hapten complex. The distances from the bound metal ion to the imidazole side chains of two of the three hlstldine residues of protein 315 have been determined. ¹H nuclear magnetic resonance has been used to study the histidine residues of the Fv fragment of protein 315. It has been shown that one of the three histidine residues (102<sub>H</sub>) is close to the combining site, but that this residue does not participate directly in binding haptens. <sup>31</sup> P nuclear magnetic resonance studies have shown the presence of a positively charged amino acid side chain near the entrance of the combining site of the Fv fragment. This residue has been identified as Arg 95<sub>L</sub>. The mode of binding of trini trophenyl derivatives to the Fv fragment has been studied by <sup>1</sup>H nuclear magnetic resonance. It is concluded that these haptens, when bound to the Kv fragment, make contacts with the same amino acid side chains as Dnp derivatives.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:453918 |
| Date | January 1979 |
| Creators | Dower, Steven K. |
| Contributors | Dwek, Raymond A. |
| Publisher | University of Oxford |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://ora.ox.ac.uk/objects/uuid:735da1be-67eb-4cb0-a774-e7343362fc8e |
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