In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes.
In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes.
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DOI：
10.3390/md18090439
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发表时间：
2020-08-24
期刊：
影响因子：
5.4
通讯作者：
Zatylny-Gaudin C
中科院分区：
文献类型：
--
作者：
Benoist L;Houyvet B;Henry J;Corre E;Zanuttini B;Zatylny-Gaudin C
乌贼血细胞是抗菌肽(AMPs)的潜在来源。为了研究对华丽弧菌的免疫应答并确定新的AMP，基于差异转录研究和使用多种工具进行深入的计算机分析，开发了一种独创的方法。从墨鱼血细胞中提取到两个新的转录本。对注释的转录本的第一次分析表明，Toll/NF-κB途径成员的存在，包括新发现的因子，如SO-TLR-h，SO-IKK-h和SO-Rel/NF-κB-h。在8个Toll/NF-κB通路成员中，有7个成员在脾弧菌攻击后表达上调。此外，参与免疫反应的免疫因子也被识别并上调。然而，基于注释或保守模式搜索没有发现AMP。因此，我们使用了几种可用的工具，如自制的软件程序PepTraq，基于差异表达和序列特征对未注释的转录本进行了深入的电子分析。最后，合成了5个AMP候选化合物。其中，NF19、AV19和GK28对革兰氏阴性菌具有一定的抗菌活性。每种多肽都有不同的活性谱，特别是对弧菌物种的活性。最有活性的多肽GK28不溶血，而NF19和AV19在50-100µM浓度范围内溶血，比其最低抑菌浓度高5-10倍。
Cuttlefish (Sepia officinalis) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to Vibrio splendidus 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 V. splendidus or not. A first analysis of the annotated transcripts revealed the presence of Toll/NF-κB pathway members, including newly identified factors such as So-TLR-h, So-IKK-h and So-Rel/NF-κB-h. Out of the eight Toll/NF-κB pathway members, seven were found up-regulated following V. splendidus 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 Vibrio 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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影响因子：
2.9
作者：
Fernandes, JMO;Molle, G;Smith, VJ
通讯作者：
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DOI：
10.1016/s1570-9639(03)00018-9
发表时间：
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