2b68: Difference between revisions

Revision as of 17:34, 21 February 2008

Solution structure of the recombinant Crassostrea gigas defensin

OverviewOverview

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-alphabeta) 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.

About this StructureAbout this Structure

2B68 is a Single protein structure of sequence from Crassostrea gigas. Full crystallographic information is available from OCA.

ReferenceReference

Characterization of a defensin from the oyster Crassostrea gigas. Recombinant production, folding, solution structure, antimicrobial activities, and gene expression., Gueguen Y, Herpin A, Aumelas A, Garnier J, Fievet J, Escoubas JM, Bulet P, Gonzalez M, Lelong C, Favrel P, Bachere E, J Biol Chem. 2006 Jan 6;281(1):313-23. Epub 2005 Oct 24. PMID:16246846

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 ==Overview==
-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-alphabeta) 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.
+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-alphabeta) 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.
 ==About this Structure==
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 [[Category: Bachere, E.]]
 [[Category: Bulet, P.]]
-[[Category: Escoubas, J.M.]]
+[[Category: Escoubas, J M.]]
 [[Category: Favrel, P.]]
 [[Category: Fievet, J.]]
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 [[Category: defensin]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jan 29 18:17:51 2008''
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