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Engineering of protein epitopes: a single deletion in a snake toxin generates full binding capacity to a previously unrecognized antibody.
- Source :
-
Protein engineering [Protein Eng] 1994 Jul; Vol. 7 (7), pp. 917-23. - Publication Year :
- 1994
-
Abstract
- Structural features associated with the ability of a monoclonal antibody (mAb) to discriminate between protein variants are identified and engineered. The variants are the curaremimetic toxin alpha from Naja nigricollis and erabutoxin a or b from Laticauda semifasciata, which differ from each other by 16 substitutions and one insertion. The neutralizing mAb M alpha 1 recognizes with high affinity a topographical epitope on the surface of toxin alpha, but fails to recognize the erabutoxins although they possess most of the residues forming the presumed epitope. Examinations of the toxin alpha and erabutoxin 3-D structures and molecular dynamics simulations reveal several differences between the variants. In particular, the region involving the beta-turn 17-24 is organized differently. Analysis of the differences found in this region suggest that the insertion (or deletion) at position 18 of the variant amino acid sequences is particularly important in determining the differential cross-reactivity. To test this proposal, residue 18 was deleted in one erabutoxin using site-directed mutagenesis, and the biological properties of the resulting mutant were examined. We found that full antigenicity was restored in the previously unrecognized variant. The implications of this finding are discussed.
- Subjects :
- Amino Acid Sequence
Antigen-Antibody Reactions
Cholinergic Antagonists
Cobra Neurotoxin Proteins chemistry
Cobra Neurotoxin Proteins genetics
Computer Simulation
Cross Reactions
Erabutoxins chemistry
Erabutoxins genetics
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
Protein Conformation
Recombinant Fusion Proteins chemistry
Sequence Deletion
Antibodies, Monoclonal immunology
Cobra Neurotoxin Proteins immunology
Epitopes chemistry
Erabutoxins immunology
Protein Engineering
Subjects
Details
- Language :
- English
- ISSN :
- 0269-2139
- Volume :
- 7
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Protein engineering
- Publication Type :
- Academic Journal
- Accession number :
- 7526378
- Full Text :
- https://doi.org/10.1093/protein/7.7.917