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An Antibody Binding Site on Cytochrome C Defined by Hydrogen Exchange and Two-Dimensional NMR
Open accessAbstract
The interaction of a protein antigen, horse cytochrome c (cyt c), with a monoclonal antibody has been studied by hydrogen-deuterium (H-D) exchange labeling and two-dimensional nuclear magnetic resonance (2D NMR) methods. The H-exchange rate of residues in three discontiguous regions of the cyt c polypeptide backbone was slowed by factors up to 340-fold in the antibody-antigen complex compared with free cyt c. The protected residues, 36 to 38, 59, 60, 64 to 67, 100, and 101, and their hydrogen-bond acceptors, are brought together in the three-dimensional structure to form a contiguous, largely exposed protein surface with an area of about 750 square angstroms. The interaction site determined in this way is consistent with prior epitope mapping studies and includes several residues that were not previously identified. The hydrogen exchange labeling approach can be used to map binding sites on small proteins in antibody-antigen complexes and may be applicable to protein-protein and protein-ligand interactions in general.
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Citations
Cited by
Zhang, Z., & Smith, D. L. (1993) · Protein Science · doi:10.1002/pro.5560020404 · Cited by 1,012
Feng, S., Chen, J. K., Yu, H., Simon, J. A., & Schreiber, S. L. (1994) · Science · doi:10.1126/science.7526465 · Cited by 687
Mechanisms and uses of hydrogen exchange
Englander, S. W., Sosnick, T. R., Englander, J. J., & Mayne, L. (1996) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(96)80090-x · Cited by 383
Self-citation[18] Systematic mutational analyses of protein-protein interfaces
(1991) · Methods in Enzymology · doi:10.1016/0076-6879(91)02020-a · Cited by 345
Pharmacological uses and perspectives of heavy water and deuterated compounds
Kushner, D. J., Baker, A., & Dunstall, T. G. (1999) · Canadian Journal of Physiology and Pharmacology · doi:10.1139/cjpp-77-2-79 · Cited by 332
Metadata
- Authors
- Paterson, Y., Englander, S. W., & Roder, H.
- Venue
- Science
- Published
- 1990-08-17
- Volume
- 249
- Issue
- 4970
- Pages
- 755-759
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2076576008
- Cited by
- 186
- References
- 0
- FWCI
- 4.19
- Type
- journal article
- Language
- en
- Open access
- green
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- Paterson, Y., Englander, S. W., & Roder, H. (1990). An Antibody Binding Site on Cytochrome C Defined by Hydrogen Exchange and Two-Dimensional NMR. Science. https://doi.org/10.1126/science.1697101
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