In-Depth In Silico Search for Cuttlefish ( Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes
Cuttlefish ( ) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to and identify new AMPs, an original approach was developed based on a differential transcriptomic study and an in-depth in silico analysis using multiple tools. Two de novo transcriptomes...
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Published in | Marine drugs Vol. 18; no. 9; p. 439 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
Switzerland
MDPI AG
24.08.2020
MDPI |
Subjects | |
Online Access | Get full text |
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Summary: | Cuttlefish (
) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to
and identify new AMPs, an original approach was developed based on a differential transcriptomic study and an in-depth in silico analysis using multiple tools. Two de novo transcriptomes were retrieved from cuttlefish haemocytes following challenge by
.
or not. A first analysis of the annotated transcripts revealed the presence of Toll/NF-κB pathway members, including newly identified factors such as
-TLR-h,
-IKK-h and
-Rel/NF-κB-h. Out of the eight Toll/NF-κB pathway members, seven were found up-regulated following
.
challenge. Besides, immune factors involved in the immune response were also identified and up-regulated. However, no AMP was identified based on annotation or conserved pattern searches. We therefore performed an in-depth in silico analysis of unannotated transcripts based on differential expression and sequence characteristics, using several tools available like PepTraq, a homemade software program. Finally, five AMP candidates were synthesized. Among them, NF19, AV19 and GK28 displayed antibacterial activity against Gram-negative bacteria. Each peptide had a different spectrum of activity, notably against
species. GK28-the most active peptide-was not haemolytic, whereas NF19 and AV19 were haemolytic at concentrations between 50 and 100 µM, 5 to 10 times higher than their minimum inhibitory concentration. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1660-3397 1660-3397 |
DOI: | 10.3390/MD18090439 |