Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution
Characterizing and understanding the antibody binding interface have become a pre-requisite for rational antibody design and engineering. The antigen-binding site is formed by six hypervariable loops, known as the complementarity determining regions (CDRs) and by the relative interdomain orientation...
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2021-03-01
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doaj-d1ec0d04e2e54bd297be3ddaffc181ec2021-03-02T05:05:43ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-03-011210.3389/fimmu.2021.630034630034Mutation of Framework Residue H71 Results in Different Antibody Paratope States in SolutionMonica L. Fernández-QuinteroKatharina B. KroellFlorian HoferJakob R. RiccabonaKlaus R. LiedlCharacterizing and understanding the antibody binding interface have become a pre-requisite for rational antibody design and engineering. The antigen-binding site is formed by six hypervariable loops, known as the complementarity determining regions (CDRs) and by the relative interdomain orientation (VH–VL). Antibody CDR loops with a certain sequence have been thought to be limited to a single static canonical conformation determining their binding properties. However, it has been shown that antibodies exist as ensembles of multiple paratope states, which are defined by a characteristic combination of CDR loop conformations and interdomain orientations. In this study, we thermodynamically and kinetically characterize the prominent role of residue 71H (Chothia nomenclature), which does not only codetermine the canonical conformation of the CDR-H2 loop but also results in changes in conformational diversity and population shifts of the CDR-H1 and CDR-H3 loop. As all CDR loop movements are correlated, conformational rearrangements of the heavy chain CDR loops also induce conformational changes in the CDR-L1, CDR-L2, and CDR-L3 loop. These overall conformational changes of the CDR loops also influence the interface angle distributions, consequentially leading to different paratope states in solution. Thus, the type of residue of 71H, either an alanine or an arginine, not only influences the CDR-H2 loop ensembles, but co-determines the paratope states in solution. Characterization of the functional consequences of mutations of residue 71H on the paratope states and interface orientations has broad implications in the field of antibody engineering.https://www.frontiersin.org/articles/10.3389/fimmu.2021.630034/fullantibodiescanonical clustersmolecular dynamics simulationsrole of residue 71HMarkov-state modes |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Monica L. Fernández-Quintero Katharina B. Kroell Florian Hofer Jakob R. Riccabona Klaus R. Liedl |
spellingShingle |
Monica L. Fernández-Quintero Katharina B. Kroell Florian Hofer Jakob R. Riccabona Klaus R. Liedl Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution Frontiers in Immunology antibodies canonical clusters molecular dynamics simulations role of residue 71H Markov-state modes |
author_facet |
Monica L. Fernández-Quintero Katharina B. Kroell Florian Hofer Jakob R. Riccabona Klaus R. Liedl |
author_sort |
Monica L. Fernández-Quintero |
title |
Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution |
title_short |
Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution |
title_full |
Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution |
title_fullStr |
Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution |
title_full_unstemmed |
Mutation of Framework Residue H71 Results in Different Antibody Paratope States in Solution |
title_sort |
mutation of framework residue h71 results in different antibody paratope states in solution |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2021-03-01 |
description |
Characterizing and understanding the antibody binding interface have become a pre-requisite for rational antibody design and engineering. The antigen-binding site is formed by six hypervariable loops, known as the complementarity determining regions (CDRs) and by the relative interdomain orientation (VH–VL). Antibody CDR loops with a certain sequence have been thought to be limited to a single static canonical conformation determining their binding properties. However, it has been shown that antibodies exist as ensembles of multiple paratope states, which are defined by a characteristic combination of CDR loop conformations and interdomain orientations. In this study, we thermodynamically and kinetically characterize the prominent role of residue 71H (Chothia nomenclature), which does not only codetermine the canonical conformation of the CDR-H2 loop but also results in changes in conformational diversity and population shifts of the CDR-H1 and CDR-H3 loop. As all CDR loop movements are correlated, conformational rearrangements of the heavy chain CDR loops also induce conformational changes in the CDR-L1, CDR-L2, and CDR-L3 loop. These overall conformational changes of the CDR loops also influence the interface angle distributions, consequentially leading to different paratope states in solution. Thus, the type of residue of 71H, either an alanine or an arginine, not only influences the CDR-H2 loop ensembles, but co-determines the paratope states in solution. Characterization of the functional consequences of mutations of residue 71H on the paratope states and interface orientations has broad implications in the field of antibody engineering. |
topic |
antibodies canonical clusters molecular dynamics simulations role of residue 71H Markov-state modes |
url |
https://www.frontiersin.org/articles/10.3389/fimmu.2021.630034/full |
work_keys_str_mv |
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