Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression
In invertebrates, defensins were found in arthropods and in the mussels. Here, we report for the first time the identification and characterization of a defensin (Cg-Def) from an oyster. Cg-def mRNA was isolated from Crassostrea gigas mantle using an expressed sequence tag approach. To gain insight...
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American Society for Biochemistry and Molecular Biology
2006
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Online Access: | https://archimer.ifremer.fr/doc/2006/publication-2127.pdf https://doi.org/10.1074/jbc.M510850200 https://archimer.ifremer.fr/doc/00000/2127/ |
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ftarchimer:oai:archimer.ifremer.fr:2127 2023-05-15T15:57:51+02:00 Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression Gueguen, Yannick Herpin, Amaury Aumelas, André Garnier, Julien Fievet, Julie Escoubas, Jean-michel Bulet, Philippe Gonzalez, Marcelo Lelong, Christophe Favrel, Pascal Bachere, Evelyne 2006 application/pdf https://archimer.ifremer.fr/doc/2006/publication-2127.pdf https://doi.org/10.1074/jbc.M510850200 https://archimer.ifremer.fr/doc/00000/2127/ eng eng American Society for Biochemistry and Molecular Biology https://archimer.ifremer.fr/doc/2006/publication-2127.pdf doi:10.1074/jbc.M510850200 https://archimer.ifremer.fr/doc/00000/2127/ 2006 American Society for Biochemistry and Molecular Biology info:eu-repo/semantics/openAccess restricted use Journal of Biological Inorganic Chemistry (0021-9258) (American Society for Biochemistry and Molecular Biology), 2006 , Vol. 281 , N. 1 , P. 313-323 Crassostrea gigas Genomic library Microbiology Defensin Oyster text Publication info:eu-repo/semantics/article 2006 ftarchimer https://doi.org/10.1074/jbc.M510850200 2021-09-23T20:14:29Z In invertebrates, defensins were found in arthropods and in the mussels. Here, we report for the first time the identification and characterization of a defensin (Cg-Def) from an oyster. Cg-def mRNA was isolated from Crassostrea gigas mantle using an expressed sequence tag approach. To gain insight into potential roles of Cg-Def in oyster immunity, we produced the recombinant peptide in Escherichia coli, characterized its antimicrobial activities, determined its solution structure by NMR spectroscopy, and quantified its gene expression in vivo following bacterial challenge of oysters. Recombinant Cg-Def was active in vitro against Gram-positive bacteria but showed no or limited activities against Gram-negative bacteria and fungi. The activity of Cg-Def was retained in vitro at a salt concentration similar to that of seawater. The Cg-Def structure shares the so-called cystine-stabilized alpha-beta motif (CS-alpha beta) with arthropod defensins but is characterized by the presence of an additional disulfide bond, as previously observed in the mussel defensin (MGD-1). Nevertheless, despite a similar global fold, the Cg-Def and MGD-1 structures mainly differ by the size of their loops and by the presence of two aspartic residues in Cg-Def. Distribution of Cg-def mRNA in various oyster tissues revealed that Cg-def is mainly expressed in mantle edge where it was detected by mass spectrometry analyses. Furthermore, we observed that the Cg-def messenger concentration was unchanged after bacterial challenge. Our results suggest that Cg-def gene is continuously expressed in the mantle and would play a key role in oyster by providing a first line of defense against pathogen colonization. Article in Journal/Newspaper Crassostrea gigas Archimer (Archive Institutionnelle de l'Ifremer - Institut français de recherche pour l'exploitation de la mer) Journal of Biological Chemistry 281 1 313 323 |
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Open Polar |
collection |
Archimer (Archive Institutionnelle de l'Ifremer - Institut français de recherche pour l'exploitation de la mer) |
op_collection_id |
ftarchimer |
language |
English |
topic |
Crassostrea gigas Genomic library Microbiology Defensin Oyster |
spellingShingle |
Crassostrea gigas Genomic library Microbiology Defensin Oyster Gueguen, Yannick Herpin, Amaury Aumelas, André Garnier, Julien Fievet, Julie Escoubas, Jean-michel Bulet, Philippe Gonzalez, Marcelo Lelong, Christophe Favrel, Pascal Bachere, Evelyne Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
topic_facet |
Crassostrea gigas Genomic library Microbiology Defensin Oyster |
description |
In invertebrates, defensins were found in arthropods and in the mussels. Here, we report for the first time the identification and characterization of a defensin (Cg-Def) from an oyster. Cg-def mRNA was isolated from Crassostrea gigas mantle using an expressed sequence tag approach. To gain insight into potential roles of Cg-Def in oyster immunity, we produced the recombinant peptide in Escherichia coli, characterized its antimicrobial activities, determined its solution structure by NMR spectroscopy, and quantified its gene expression in vivo following bacterial challenge of oysters. Recombinant Cg-Def was active in vitro against Gram-positive bacteria but showed no or limited activities against Gram-negative bacteria and fungi. The activity of Cg-Def was retained in vitro at a salt concentration similar to that of seawater. The Cg-Def structure shares the so-called cystine-stabilized alpha-beta motif (CS-alpha beta) with arthropod defensins but is characterized by the presence of an additional disulfide bond, as previously observed in the mussel defensin (MGD-1). Nevertheless, despite a similar global fold, the Cg-Def and MGD-1 structures mainly differ by the size of their loops and by the presence of two aspartic residues in Cg-Def. Distribution of Cg-def mRNA in various oyster tissues revealed that Cg-def is mainly expressed in mantle edge where it was detected by mass spectrometry analyses. Furthermore, we observed that the Cg-def messenger concentration was unchanged after bacterial challenge. Our results suggest that Cg-def gene is continuously expressed in the mantle and would play a key role in oyster by providing a first line of defense against pathogen colonization. |
format |
Article in Journal/Newspaper |
author |
Gueguen, Yannick Herpin, Amaury Aumelas, André Garnier, Julien Fievet, Julie Escoubas, Jean-michel Bulet, Philippe Gonzalez, Marcelo Lelong, Christophe Favrel, Pascal Bachere, Evelyne |
author_facet |
Gueguen, Yannick Herpin, Amaury Aumelas, André Garnier, Julien Fievet, Julie Escoubas, Jean-michel Bulet, Philippe Gonzalez, Marcelo Lelong, Christophe Favrel, Pascal Bachere, Evelyne |
author_sort |
Gueguen, Yannick |
title |
Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
title_short |
Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
title_full |
Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
title_fullStr |
Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
title_full_unstemmed |
Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
title_sort |
characterization of a defensin from the oyster crassostrea gigas - recombinant production, folding, solution structure, antimicrobial activities, and gene expression |
publisher |
American Society for Biochemistry and Molecular Biology |
publishDate |
2006 |
url |
https://archimer.ifremer.fr/doc/2006/publication-2127.pdf https://doi.org/10.1074/jbc.M510850200 https://archimer.ifremer.fr/doc/00000/2127/ |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_source |
Journal of Biological Inorganic Chemistry (0021-9258) (American Society for Biochemistry and Molecular Biology), 2006 , Vol. 281 , N. 1 , P. 313-323 |
op_relation |
https://archimer.ifremer.fr/doc/2006/publication-2127.pdf doi:10.1074/jbc.M510850200 https://archimer.ifremer.fr/doc/00000/2127/ |
op_rights |
2006 American Society for Biochemistry and Molecular Biology info:eu-repo/semantics/openAccess restricted use |
op_doi |
https://doi.org/10.1074/jbc.M510850200 |
container_title |
Journal of Biological Chemistry |
container_volume |
281 |
container_issue |
1 |
container_start_page |
313 |
op_container_end_page |
323 |
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1766393558441394176 |