Mapping of IFN-beta epitopes important for receptor binding and biologic activation: comparison of results achieved using antibody-based methods and alanine substitution mutagenesis
| Autor: | Darren P. Baker, Carole de Dios, Zhifang Li, Celine Muldowney, Paula S. Hochman, Matthew Betzenhauser, Sidney E. Grossberg, Miller Stephan S, Philip N. Redlich, Laura Runkel, Michael Karpusas, Adrian Whitty, Mohammad Zafari |
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| Rok vydání: | 2001 |
| Předmět: |
Models
Molecular medicine.drug_class Immunology Molecular Sequence Data Receptor Interferon alpha-beta Monoclonal antibody Binding Competitive Epitope Epitopes Antibody Specificity Neutralization Tests Virology medicine Humans Amino Acid Sequence Binding site Receptor Receptors Interferon Alanine Dose-Response Relationship Drug Chemistry Mutagenesis Antibodies Monoclonal Membrane Proteins Cell Biology Interferon-beta Alanine scanning Molecular biology Epitope mapping Biochemistry Amino Acid Substitution Sequence Alignment Epitope Mapping Interferon beta-1a |
| Zdroj: | Journal of interferoncytokine research : the official journal of the International Society for Interferon and Cytokine Research. 21(11) |
| ISSN: | 1079-9907 |
| Popis: | The epitopes important for receptor binding and activation of human interferon-beta1a (IFN-beta1a) were mapped with monoclonal antibodies (mAb), grouped on the basis of their specificity and ability to neutralize biologic activity, and alanine scanning mutagenesis (ASM). The binding properties of nine mAb were defined, using ASM-IFN-beta mutants having alanine substituted at targeted, surface-exposed residues. The results were correlated with the mAb neutralizing potency. Of six mAb that bound either at or adjacent to the IFNAR-2 receptor chain binding site defined by the ASM epitopes, only three had measurable neutralizing activity. Two of these inhibited IFN-beta/IFNAR-2 complex formation, suggesting that steric hindrance of receptor binding constitutes their mechanism of neutralization. However, two mAb that bound to sites remote from the IFNAR-2 binding site on IFN-beta also inhibited IFN-beta/IFNAR-2 complex formation and demonstrated potent neutralizing activity. Thus, neutralizing mAb may employ mechanisms other than steric blockade to inhibit directly the binding of receptor by cytokine, limiting their usefulness as tools to define precise receptor-ligand interaction sites. |
| Databáze: | OpenAIRE |
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