Functional Mapping of Conserved Residues Located at the VL and VH Domain Interface of a Fab
| العنوان: | Functional Mapping of Conserved Residues Located at the VL and VH Domain Interface of a Fab |
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| المؤلفون: | Danièle Altschuh, Jean Chatellier, Marc H.V. Van Regenmortel, Thierry Vernet |
| المساهمون: | Laboratoire d'Ingénierie des Macromolécules (LIM), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de biologie structurale (IBS - UMR 5075), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS) |
| المصدر: | Journal of Molecular Biology Journal of Molecular Biology, Elsevier, 1996, 264 (1), pp.1-6. ⟨10.1006/jmbi.1996.0618⟩ Journal of Molecular Biology, 1996, 264 (1), pp.1-6. ⟨10.1006/jmbi.1996.0618⟩ |
| بيانات النشر: | Elsevier BV, 1996. |
| سنة النشر: | 1996 |
| مصطلحات موضوعية: | MESH: Immunoglobulin Light Chains, Mutant, Immunoglobulin Variable Region, MESH: Immunoglobulin Variable Region, MESH: Protein Structure, Secondary, Peptide, MESH: Amino Acid Sequence, Antibodies, Viral, MESH: Peptide Mapping, Protein Structure, Secondary, Antigen-Antibody Reactions, Mice, 0302 clinical medicine, Structural Biology, MESH: Antigen-Antibody Reactions, MESH: Capsid, MESH: Animals, MESH: Peptide Fragments, Conserved Sequence, Alanine, chemistry.chemical_classification, 0303 health sciences, MESH: Conserved Sequence, MESH: Kinetics, biology, Chemistry, Alanine scanning, MESH: Binding Sites, Antibody, Receptor–ligand kinetics, 3. Good health, Tobacco Mosaic Virus, MESH: Immunoglobulin Fab Fragments, MESH: Mutagenesis, Site-Directed, Biochemistry, 030220 oncology & carcinogenesis, Antibody, Immunoglobulin Heavy Chains, MESH: Immunoglobulin Heavy Chains, Mutagenesis (molecular biology technique), Peptide Mapping, Immunoglobulin Fab Fragments, 03 medical and health sciences, Capsid, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, MESH: Mice, Molecular Biology, 030304 developmental biology, Peptide Fragments, Kinetics, Mutagenesis, Site-Directed, biology.protein, Immunoglobulin Light Chains, Paratope, Binding Sites, Antibody, MESH: Tobacco Mosaic Virus, MESH: Antibodies, Viral |
| الوصف: | The interface between the VL and VH domains of antibodies is highly conserved. To investigate the influence of conserved interface residues on Fab function, 13 interface residues were subjected to codon-based combinatorial alanine scanning mutagenesis in Fab 57P, specific for peptide 134 to 151 of the coat protein of tobacco mosaic virus. The 13 single mutants were analysed by Western blot to determine the effect of interface modifications on Fab expression. The kinetic rate constants of peptide-Fab mutant interactions were measured using the biosensor technology. Alanine replacements did not prevent assembly of the mutated Fabs and led to a modification of their binding properties in every case. Twelve of the 13 target residues correspond to homologous positions in the VL and VH domains, which have similar folds. Mutation at homologous positions mostly had different effects on antigen binding affinity. The replacement of bulky side-chains had the most drastic effect on binding. When smaller side-chains were replaced by alanine, the binding properties of Fab mutants differed slightly (by less than a factor of two), but significantly from that of Fab 57P. Modification of some of these residues, which are located 9 to 12 A away from the base of CDR loops, is unlikely to alter loop conformation. They may affect antigen binding indirectly by influencing the relative position of the VL and VH domains. Our results demonstrate that residues situated at the VL – VH interface and which are remote from the paratope are able to influence the antigen binding properties of antibodies. |
| تدمد: | 0022-2836 1089-8638 |
| DOI: | 10.1006/jmbi.1996.0618 |
| DOI: | 10.1006/jmbi.1996.0618⟩ |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::391a5349f046f8148eca2aae5e1d8683 https://doi.org/10.1006/jmbi.1996.0618 |
| Rights: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....391a5349f046f8148eca2aae5e1d8683 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00222836 10898638 |
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| DOI: | 10.1006/jmbi.1996.0618 |