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Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation

Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes...

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Autores principales: Dixon, Emma V., Claridge, Jolyon K., Harvey, David J., Baruah, Kavitha, Yu, Xiaojie, Vesiljevic, Snezana, Mattick, Susan, Pritchard, Laura K., Krishna, Benjamin, Scanlan, Christopher N., Schnell, Jason R., Higgins, Matthew K., Zitzmann, Nicole, Crispin, Max
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4022860/
https://www.ncbi.nlm.nih.gov/pubmed/24668806
http://dx.doi.org/10.1074/jbc.M113.532812
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author Dixon, Emma V.
Claridge, Jolyon K.
Harvey, David J.
Baruah, Kavitha
Yu, Xiaojie
Vesiljevic, Snezana
Mattick, Susan
Pritchard, Laura K.
Krishna, Benjamin
Scanlan, Christopher N.
Schnell, Jason R.
Higgins, Matthew K.
Zitzmann, Nicole
Crispin, Max
author_facet Dixon, Emma V.
Claridge, Jolyon K.
Harvey, David J.
Baruah, Kavitha
Yu, Xiaojie
Vesiljevic, Snezana
Mattick, Susan
Pritchard, Laura K.
Krishna, Benjamin
Scanlan, Christopher N.
Schnell, Jason R.
Higgins, Matthew K.
Zitzmann, Nicole
Crispin, Max
author_sort Dixon, Emma V.
collection PubMed
description Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. Here, we investigate the mechanism of human serum IgG deactivation by EndoS. We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. Similarly, we sought to investigate which domains within EndoS could contribute to activity. Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans.
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spelling pubmed-40228602014-05-22 Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation Dixon, Emma V. Claridge, Jolyon K. Harvey, David J. Baruah, Kavitha Yu, Xiaojie Vesiljevic, Snezana Mattick, Susan Pritchard, Laura K. Krishna, Benjamin Scanlan, Christopher N. Schnell, Jason R. Higgins, Matthew K. Zitzmann, Nicole Crispin, Max J Biol Chem Glycobiology and Extracellular Matrices Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. Here, we investigate the mechanism of human serum IgG deactivation by EndoS. We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. Similarly, we sought to investigate which domains within EndoS could contribute to activity. Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. American Society for Biochemistry and Molecular Biology 2014-05-16 2014-03-25 /pmc/articles/PMC4022860/ /pubmed/24668806 http://dx.doi.org/10.1074/jbc.M113.532812 Text en © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/) applies to Author Choice Articles
spellingShingle Glycobiology and Extracellular Matrices
Dixon, Emma V.
Claridge, Jolyon K.
Harvey, David J.
Baruah, Kavitha
Yu, Xiaojie
Vesiljevic, Snezana
Mattick, Susan
Pritchard, Laura K.
Krishna, Benjamin
Scanlan, Christopher N.
Schnell, Jason R.
Higgins, Matthew K.
Zitzmann, Nicole
Crispin, Max
Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation
title Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation
title_full Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation
title_fullStr Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation
title_full_unstemmed Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation
title_short Fragments of Bacterial Endoglycosidase S and Immunoglobulin G Reveal Subdomains of Each That Contribute to Deglycosylation
title_sort fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation
topic Glycobiology and Extracellular Matrices
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4022860/
https://www.ncbi.nlm.nih.gov/pubmed/24668806
http://dx.doi.org/10.1074/jbc.M113.532812
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