Specificities of Germ Line Antibodies

  • Thomas P. Theriault
  • Gordon S. Rule
  • Harden M. McConnell
Part of the NATO ASI Series book series (NSSA, volume 183)


The past few years have seen remarkable progress in research on the structure and function of antibodies. In our own work we have shown that it is possible to obtain extensive significant information on the composition and structure of antibody combining sites using NMR, together with nitroxide spin-label haptens (Anglister et al, 1984a, 1985 and 1987; Frey et al, 1984). This derived information includes the amino acid composition of the combining site region, that is, the number of tyrosines, alanines, etc. that are within ~ 20 Å of the odd electron on the paramagnetic hapten. In antibody molecules there are typically 40–50 amino acids in this combining site region. We have shown that NMR titration data can be used to estimate distances between individual protons on amino acid side chains and the odd electron (Anglister et.al., 1984b; Frey et. al., 1988). These measured distances extend out to about 20 Å, and in to distances of the order of 3–5 Å. Shorter distances can sometimes be estimated from nuclear magnetization transfer experiments. The NMR data also provide a powerful and convenient means of obtaining the on-off kinetics of hapten-antibody reactions, using resonance signals from the hapten as well as from the protein.


Light Chain Heavy Chain Germline Gene Diversity Segment XbaI Fragment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. Amit, A. G., Mariuzza, R. A., Phillips, E. V. and Poljak, R. J., 1986, Three-dimentional structure of an antigen-antibody complex at 2.8 Å resolution, Science 233:747.ADSCrossRefGoogle Scholar
  2. Anglister, J., Frey, T. and McConnell, H. M., 1984, Magnetic resonance of a monoclonal anti-spin-label antibody, Biochemistry 23:1138.CrossRefGoogle Scholar
  3. Anglister, J., Frey, T. and McConnell, H. M., 1984, Distances of tyrosine residues from a spin-label hapten in the combining site of a specific monoclonal antibody, Biochemistry 23:5372.CrossRefGoogle Scholar
  4. Anglister, J., Bond, B. W., Frey, T., Leahy, D. J., Levitt, M., McConnell, H. M., Rule, G. S., Tomasello, J. and Whittaker, M. M., 1987, Contribution of tryptophan residues to the combining site of a monoclonal anti-dinitrophenyl spin-label antibody, Biochemistry 26:6058.CrossRefGoogle Scholar
  5. Anglister, J., Frey, T. and McConnell, H. M., 1985, NMR technique for assessing contributions of heavy and light chains to an antibody combining site, Nature 315.6014:65.Google Scholar
  6. Dzierzak, E. A., Janeway, C. A., Jr., Richard, N. and Bothwell, A., 1986, Molecular characterization of antibodies bearing Id-460, J. Immunol. 136.5:1864.Google Scholar
  7. Frey, T., Anglister, J. and McConnell, H. M., 1984, Nonaromatic amino acids in the combining site of a monoclonal anti-spin-label antibody, Biochemistry 23:6470.CrossRefGoogle Scholar
  8. Frey, T., Anglister, J. and McConnell, H. M., 1988, Line-shape analysis of NMR difference spectra of an anti-spin-label antibody, Biochemistry 27:5161.CrossRefGoogle Scholar
  9. Gearhart, P. J. and Brogenhagen, D. F., 1983, Clusters of point mutations are found exclusively around rearranged antibody variable genes, Proc. Natl. Acad. Sci. USA 80:3439.ADSCrossRefGoogle Scholar
  10. Leahy, D. J., Rule, G. S., Whittaker, M. M. and McConnell, H. M., 1988, Sequences of 12 monoclonal anti-dinitrophenyl spin-label antibodies for NMR studies, Proc. Natl. Acad. Sci. USA 85:3661.ADSCrossRefGoogle Scholar
  11. Kabat, E. A., Wu, T. T., Reid-Miller, M., Perry, H. M. and Gottesman, K. S., 1987, “Sequences of Proteins of Immunological Interest”, Natl. Inst, of Health, Bethesda, MD.Google Scholar
  12. Maniatis, T., Fritsch, E. F. and Sambrook, J., 1982, “Molecular Cloning: A Laboratory Manual”, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  13. Sheriff, S., Silverton, E. W., Padlan, E. A., Cohen, G. H., Smith-Gill, S. J., Finzel, B.C. and Davies, D. R., 1987, Three-dimensional structure of an antibody-antigen complex, Proc. Natl. Acad. Sci. USA 84:8075.ADSCrossRefGoogle Scholar
  14. Southern, E., 1975, Detection of specific sequences among DNA fragments separated by gel electrophoresis, J. Mol. Biol. 98:503.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Thomas P. Theriault
    • 1
  • Gordon S. Rule
    • 1
  • Harden M. McConnell
    • 1
  1. 1.Stauffer Laboratory for Physical Chemistry, Department of ChemistryStanford UniversityStanfordUSA

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