Contribution of Tryptophan Residues to the Combining Site of a Monoclonal Anti Dinitrophenyl Spin-Label Antibody
Abstract
Two Fab fragments of the monoclonal anti dinitrophenyl (DNP) spin- label antibody AN02 were prepared by recombination of specifically deuteriated heavy and light chains. In the recombinant H(I)L(II) all the tyrosines and phenylalanines were perdeuteriated as were the tryptophan residues of the heavy chain. In the recombinant H(II)L(I) all the tyrosines and phenylalanines were perdeuteriated as were the tryptophan residues of the light chain. Saturation of three resonances of H(I)L(II), assigned to tryptophan protons of the light chain, resulted in magnetization transfer to the aromatic proton at position 6 of the DNP ring and to the CH2 protons of the glycines linked to the DNP in a diamagnetic hapten (DNP-DG). Saturation of three resonances of H(II)L(I) assigned to tryptophan protons of the heavy chain resulted in magnetization transfer to the CH2 protons of the glycines in DNP-DG. From the dependence of the magnetization transfer on the irradiation time, the cross relaxation rates between the involved protons were estimated. The inferred distances between these protons of the hapten and certain tryptophan protons are 3-4 A. It is concluded that in the combining site of AN02 there is one tryptophan from the light chain and one tryptophan from the heavy chain that are very near the hapten, identified as tryptophan-91 of the light chain and tryptophan-47 of the heavy chain. Reprints. (EDC)
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1987
- Accession Number
- ADA219760
Entities
People
- Daniel Leahy
- Jacob Anglister
- Martha W. Bond
- Michael Levitt
- Tom Frey
Organizations
- Stanford University