Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope
Conformational rearrangements in antibody antigen recognition are essential events where kinetic discrimination of isomers expands the universe of combinations. We investigated the interaction mechanism of a monoclonal antibody, M1, raised against E7 from human papillomavirus, a prototypic viral onc...
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2013
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v288_n18_p13110_Fassolari http://hdl.handle.net/20.500.12110/paper_00219258_v288_n18_p13110_Fassolari |
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paper:paper_00219258_v288_n18_p13110_Fassolari2023-06-08T14:43:33Z Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope Antigen presenting cells Antigen-antibody interaction High-energy barriers Immunodominant epitopes Interaction mechanisms Intrinsically disordered proteins Intrinsically disordered regions Mutagenesis experiment Amino acids Antigen-antibody reactions Association reactions Clone cells Experiments Isomerization Isomers Kinetics Monoclonal antibodies Epitopes amino acid epitope isoprotein monoclonal antibody oncoprotein proline proline derivative virus protein amino terminal sequence antibody affinity antibody specificity antigen antibody reaction antigen presentation article carboxy terminal sequence isomerization kinetics mutagenesis priority journal protein conformation protein defect protein domain protein secondary structure Wart virus Antigen Antigen Recognition Biophysics Conformational Selection E7 Oncoprotein Intrinsically Disordered Proteins Pre-steady-state Kinetics Prolyl Isomerization Protein Folding Animals Antibodies, Monoclonal, Murine-Derived Antibodies, Viral Antibody Specificity Epitopes Human papillomavirus 16 Humans Mice Nuclear Magnetic Resonance, Biomolecular Papillomavirus E7 Proteins Protein Structure, Secondary Human papillomavirus Conformational rearrangements in antibody antigen recognition are essential events where kinetic discrimination of isomers expands the universe of combinations. We investigated the interaction mechanism of a monoclonal antibody, M1, raised against E7 from human papillomavirus, a prototypic viral oncoprotein and a model intrinsically disordered protein. The mapped 12-amino acid immunodominant epitope lies within a "hinge" region between the N-terminal intrinsically disordered and the C-terminal globular domains. Kinetic experiments show that despite being within an intrinsically disordered region, the hinge E7 epitope has at least two populations separated by a high energy barrier. Nuclear magnetic resonance traced the origin of this barrier to a very slow (t1/2 ∼4 min) transcis prolyl isomerization event involving changes in secondary structure. The less populated (10%) cis isomer is the binding-competent species, thus requiring the 90% of molecules in the trans configuration to isomerize before binding. The association rate for the cis isomer approaches 6 × 107 M-1 s-1, a ceiling for antigen-antibody interactions. Mutagenesis experiments showed that Pro-41 in E7Ep was required for both binding and isomerization. After a slow postbinding unimolecular rearrangement, a consolidated complex with K D = 1.2 × 10-7 M is reached. Our results suggest that presentation of this viral epitope by the antigen-presenting cells would have to be "locked" in the cis conformation, in opposition to the most populated trans isomer, in order to select the specific antibody clone that goes through affinity and kinetic maturation. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc. 2013 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v288_n18_p13110_Fassolari http://hdl.handle.net/20.500.12110/paper_00219258_v288_n18_p13110_Fassolari |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Antigen presenting cells Antigen-antibody interaction High-energy barriers Immunodominant epitopes Interaction mechanisms Intrinsically disordered proteins Intrinsically disordered regions Mutagenesis experiment Amino acids Antigen-antibody reactions Association reactions Clone cells Experiments Isomerization Isomers Kinetics Monoclonal antibodies Epitopes amino acid epitope isoprotein monoclonal antibody oncoprotein proline proline derivative virus protein amino terminal sequence antibody affinity antibody specificity antigen antibody reaction antigen presentation article carboxy terminal sequence isomerization kinetics mutagenesis priority journal protein conformation protein defect protein domain protein secondary structure Wart virus Antigen Antigen Recognition Biophysics Conformational Selection E7 Oncoprotein Intrinsically Disordered Proteins Pre-steady-state Kinetics Prolyl Isomerization Protein Folding Animals Antibodies, Monoclonal, Murine-Derived Antibodies, Viral Antibody Specificity Epitopes Human papillomavirus 16 Humans Mice Nuclear Magnetic Resonance, Biomolecular Papillomavirus E7 Proteins Protein Structure, Secondary Human papillomavirus |
| spellingShingle |
Antigen presenting cells Antigen-antibody interaction High-energy barriers Immunodominant epitopes Interaction mechanisms Intrinsically disordered proteins Intrinsically disordered regions Mutagenesis experiment Amino acids Antigen-antibody reactions Association reactions Clone cells Experiments Isomerization Isomers Kinetics Monoclonal antibodies Epitopes amino acid epitope isoprotein monoclonal antibody oncoprotein proline proline derivative virus protein amino terminal sequence antibody affinity antibody specificity antigen antibody reaction antigen presentation article carboxy terminal sequence isomerization kinetics mutagenesis priority journal protein conformation protein defect protein domain protein secondary structure Wart virus Antigen Antigen Recognition Biophysics Conformational Selection E7 Oncoprotein Intrinsically Disordered Proteins Pre-steady-state Kinetics Prolyl Isomerization Protein Folding Animals Antibodies, Monoclonal, Murine-Derived Antibodies, Viral Antibody Specificity Epitopes Human papillomavirus 16 Humans Mice Nuclear Magnetic Resonance, Biomolecular Papillomavirus E7 Proteins Protein Structure, Secondary Human papillomavirus Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| topic_facet |
Antigen presenting cells Antigen-antibody interaction High-energy barriers Immunodominant epitopes Interaction mechanisms Intrinsically disordered proteins Intrinsically disordered regions Mutagenesis experiment Amino acids Antigen-antibody reactions Association reactions Clone cells Experiments Isomerization Isomers Kinetics Monoclonal antibodies Epitopes amino acid epitope isoprotein monoclonal antibody oncoprotein proline proline derivative virus protein amino terminal sequence antibody affinity antibody specificity antigen antibody reaction antigen presentation article carboxy terminal sequence isomerization kinetics mutagenesis priority journal protein conformation protein defect protein domain protein secondary structure Wart virus Antigen Antigen Recognition Biophysics Conformational Selection E7 Oncoprotein Intrinsically Disordered Proteins Pre-steady-state Kinetics Prolyl Isomerization Protein Folding Animals Antibodies, Monoclonal, Murine-Derived Antibodies, Viral Antibody Specificity Epitopes Human papillomavirus 16 Humans Mice Nuclear Magnetic Resonance, Biomolecular Papillomavirus E7 Proteins Protein Structure, Secondary Human papillomavirus |
| description |
Conformational rearrangements in antibody antigen recognition are essential events where kinetic discrimination of isomers expands the universe of combinations. We investigated the interaction mechanism of a monoclonal antibody, M1, raised against E7 from human papillomavirus, a prototypic viral oncoprotein and a model intrinsically disordered protein. The mapped 12-amino acid immunodominant epitope lies within a "hinge" region between the N-terminal intrinsically disordered and the C-terminal globular domains. Kinetic experiments show that despite being within an intrinsically disordered region, the hinge E7 epitope has at least two populations separated by a high energy barrier. Nuclear magnetic resonance traced the origin of this barrier to a very slow (t1/2 ∼4 min) transcis prolyl isomerization event involving changes in secondary structure. The less populated (10%) cis isomer is the binding-competent species, thus requiring the 90% of molecules in the trans configuration to isomerize before binding. The association rate for the cis isomer approaches 6 × 107 M-1 s-1, a ceiling for antigen-antibody interactions. Mutagenesis experiments showed that Pro-41 in E7Ep was required for both binding and isomerization. After a slow postbinding unimolecular rearrangement, a consolidated complex with K D = 1.2 × 10-7 M is reached. Our results suggest that presentation of this viral epitope by the antigen-presenting cells would have to be "locked" in the cis conformation, in opposition to the most populated trans isomer, in order to select the specific antibody clone that goes through affinity and kinetic maturation. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc. |
| title |
Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| title_short |
Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| title_full |
Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| title_fullStr |
Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| title_full_unstemmed |
Minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| title_sort |
minute time scale prolyl isomerization governs antibody recognition of an intrinsically disordered immunodominant epitope |
| publishDate |
2013 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v288_n18_p13110_Fassolari http://hdl.handle.net/20.500.12110/paper_00219258_v288_n18_p13110_Fassolari |
| _version_ |
1768542348651790336 |