An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody

ABSTRACT The structural and functional significance of somatic insertions and deletions in antibody chains is unclear. Here, we demonstrate that a naturally occurring three-amino-acid insertion within the influenza virus-specific human monoclonal antibody 2D1 heavy-chain variable region reconfigures...

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Autores principales:	Jens C. Krause, Damian C. Ekiert, Terrence M. Tumpey, Patricia B. Smith, Ian A. Wilson, James E. Crowe
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2011
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/048e786b5ef74f1bacc5e4ca0983aec7
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spelling	oai:doaj.org-article:048e786b5ef74f1bacc5e4ca0983aec72021-11-15T15:38:46ZAn Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody10.1128/mBio.00345-102150-7511https://doaj.org/article/048e786b5ef74f1bacc5e4ca0983aec72011-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00345-10https://doaj.org/toc/2150-7511ABSTRACT The structural and functional significance of somatic insertions and deletions in antibody chains is unclear. Here, we demonstrate that a naturally occurring three-amino-acid insertion within the influenza virus-specific human monoclonal antibody 2D1 heavy-chain variable region reconfigures the antibody-combining site and contributes to its high potency against the 1918 and 2009 pandemic H1N1 influenza viruses. The insertion arose through a series of events, including a somatic point mutation in a predicted hot-spot motif, introduction of a new hot-spot motif, a molecular duplication due to polymerase slippage, a deletion due to misalignment, and additional somatic point mutations. Atomic resolution structures of the wild-type antibody and a variant in which the insertion was removed revealed that the three-amino-acid insertion near the base of heavy-chain complementarity-determining region (CDR) H2 resulted in a bulge in that loop. This enlarged CDR H2 loop impinges on adjacent regions, causing distortion of the CDR H1 architecture and its displacement away from the antigen-combining site. Removal of the insertion restores the canonical structure of CDR H1 and CDR H2, but binding, neutralization activity, and in vivo activity were reduced markedly because of steric conflict of CDR H1 with the hemagglutinin antigen. IMPORTANCE Antibody diversification through VDJ gene recombination, junctional variation, and somatic hypermutation has clear importance for the generation of mature, high-affinity antibodies. Between 1.3 and 6.5% of antibody variable gene sequences have been reported to contain insertions or deletions, but their structural and functional significance remains less well defined. The pandemic influenza virus hemagglutinin antibody 2D1 data suggest that somatic insertions and deletions in antibody genes contribute important structural and functional features. We predict that such features can be critical for affinity and functional maturation of the human antibody repertoire.Jens C. KrauseDamian C. EkiertTerrence M. TumpeyPatricia B. SmithIan A. WilsonJames E. CroweAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 2, Iss 1 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Jens C. Krause Damian C. Ekiert Terrence M. Tumpey Patricia B. Smith Ian A. Wilson James E. Crowe An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody
description	ABSTRACT The structural and functional significance of somatic insertions and deletions in antibody chains is unclear. Here, we demonstrate that a naturally occurring three-amino-acid insertion within the influenza virus-specific human monoclonal antibody 2D1 heavy-chain variable region reconfigures the antibody-combining site and contributes to its high potency against the 1918 and 2009 pandemic H1N1 influenza viruses. The insertion arose through a series of events, including a somatic point mutation in a predicted hot-spot motif, introduction of a new hot-spot motif, a molecular duplication due to polymerase slippage, a deletion due to misalignment, and additional somatic point mutations. Atomic resolution structures of the wild-type antibody and a variant in which the insertion was removed revealed that the three-amino-acid insertion near the base of heavy-chain complementarity-determining region (CDR) H2 resulted in a bulge in that loop. This enlarged CDR H2 loop impinges on adjacent regions, causing distortion of the CDR H1 architecture and its displacement away from the antigen-combining site. Removal of the insertion restores the canonical structure of CDR H1 and CDR H2, but binding, neutralization activity, and in vivo activity were reduced markedly because of steric conflict of CDR H1 with the hemagglutinin antigen. IMPORTANCE Antibody diversification through VDJ gene recombination, junctional variation, and somatic hypermutation has clear importance for the generation of mature, high-affinity antibodies. Between 1.3 and 6.5% of antibody variable gene sequences have been reported to contain insertions or deletions, but their structural and functional significance remains less well defined. The pandemic influenza virus hemagglutinin antibody 2D1 data suggest that somatic insertions and deletions in antibody genes contribute important structural and functional features. We predict that such features can be critical for affinity and functional maturation of the human antibody repertoire.
format	article
author	Jens C. Krause Damian C. Ekiert Terrence M. Tumpey Patricia B. Smith Ian A. Wilson James E. Crowe
author_facet	Jens C. Krause Damian C. Ekiert Terrence M. Tumpey Patricia B. Smith Ian A. Wilson James E. Crowe
author_sort	Jens C. Krause
title	An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody
title_short	An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody
title_full	An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody
title_fullStr	An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody
title_full_unstemmed	An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody
title_sort	insertion mutation that distorts antibody binding site architecture enhances function of a human antibody
publisher	American Society for Microbiology
publishDate	2011
url	https://doaj.org/article/048e786b5ef74f1bacc5e4ca0983aec7
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An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody

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