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An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein

A monoclonal antibody (mAb) that binds to a transient intermediate may act as a catalyst for the corresponding reaction; here we show this principle can extend on a macro molecular scale to the induction of mutant-like oligomerization in a wild-type protein. Using the common pathogenic E342K (Z) var...

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Autores principales: Irving, James A., Miranda, Elena, Haq, Imran, Perez, Juan, Kotov, Vadim R., Faull, Sarah V., Motamedi-Shad, Neda, Lomas, David A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422257/
https://www.ncbi.nlm.nih.gov/pubmed/25738741
http://dx.doi.org/10.1042/BJ20141569
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author Irving, James A.
Miranda, Elena
Haq, Imran
Perez, Juan
Kotov, Vadim R.
Faull, Sarah V.
Motamedi-Shad, Neda
Lomas, David A.
author_facet Irving, James A.
Miranda, Elena
Haq, Imran
Perez, Juan
Kotov, Vadim R.
Faull, Sarah V.
Motamedi-Shad, Neda
Lomas, David A.
author_sort Irving, James A.
collection PubMed
description A monoclonal antibody (mAb) that binds to a transient intermediate may act as a catalyst for the corresponding reaction; here we show this principle can extend on a macro molecular scale to the induction of mutant-like oligomerization in a wild-type protein. Using the common pathogenic E342K (Z) variant of α(1)-antitrypsin as antigen–whose native state is susceptible to the formation of a proto-oligomeric intermediate–we have produced a mAb (5E3) that increases the rate of oligomerization of the wild-type (M) variant. Employing ELISA, gel shift, thermal stability and FRET time-course experiments, we show that mAb(5E3) does not bind to the native state of α(1)-antitrypsin, but recognizes a cryptic epitope in the vicinity of the post-helix A loop and strand 4C that is revealed upon transition to the polymerization intermediate, and which persists in the ensuing oligomer. This epitope is not shared by loop-inserted monomeric conformations. We show the increased amenity to polymerization by either the pathogenic E342K mutation or the binding of mAb(5E3) occurs without affecting the energetic barrier to polymerization. As mAb(5E3) also does not alter the relative stability of the monomer to intermediate, it acts in a manner similar to the E342K mutant, by facilitating the conformational interchange between these two states.
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spelling pubmed-44222572015-05-18 An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein Irving, James A. Miranda, Elena Haq, Imran Perez, Juan Kotov, Vadim R. Faull, Sarah V. Motamedi-Shad, Neda Lomas, David A. Biochem J Research Article A monoclonal antibody (mAb) that binds to a transient intermediate may act as a catalyst for the corresponding reaction; here we show this principle can extend on a macro molecular scale to the induction of mutant-like oligomerization in a wild-type protein. Using the common pathogenic E342K (Z) variant of α(1)-antitrypsin as antigen–whose native state is susceptible to the formation of a proto-oligomeric intermediate–we have produced a mAb (5E3) that increases the rate of oligomerization of the wild-type (M) variant. Employing ELISA, gel shift, thermal stability and FRET time-course experiments, we show that mAb(5E3) does not bind to the native state of α(1)-antitrypsin, but recognizes a cryptic epitope in the vicinity of the post-helix A loop and strand 4C that is revealed upon transition to the polymerization intermediate, and which persists in the ensuing oligomer. This epitope is not shared by loop-inserted monomeric conformations. We show the increased amenity to polymerization by either the pathogenic E342K mutation or the binding of mAb(5E3) occurs without affecting the energetic barrier to polymerization. As mAb(5E3) also does not alter the relative stability of the monomer to intermediate, it acts in a manner similar to the E342K mutant, by facilitating the conformational interchange between these two states. Portland Press Ltd. 2015-05-05 2015-05-15 /pmc/articles/PMC4422257/ /pubmed/25738741 http://dx.doi.org/10.1042/BJ20141569 Text en © 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY)(http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Irving, James A.
Miranda, Elena
Haq, Imran
Perez, Juan
Kotov, Vadim R.
Faull, Sarah V.
Motamedi-Shad, Neda
Lomas, David A.
An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
title An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
title_full An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
title_fullStr An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
title_full_unstemmed An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
title_short An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
title_sort antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422257/
https://www.ncbi.nlm.nih.gov/pubmed/25738741
http://dx.doi.org/10.1042/BJ20141569
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