Department of Genomic Science, School of Biological Sciences, Central University of Kerala, Kasaragod, Kerala, India.
Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia.
Sci Rep. 2023 Oct 31;13(1):18728. doi: 10.1038/s41598-023-45875-w.
Multidrug resistance has become a global health problem associated with high morbidity and mortality. Antimicrobial peptides have been acknowledged as potential leads for prospective anti-infectives. Owing to their scavenging lifestyle, Corvus splendens is thought to have developed robust immunity to pathogens found in their diet, implying that they have evolved mechanisms to resist infection. In the current study, the transcriptome of C. splendens was sequenced, and de novo assembled to identify the presence of antimicrobial peptide genes. 72.09 million high-quality clean reads were obtained which were then de novo assembled into 3,43,503 transcripts and 74,958 unigenes. About 37,559 unigenes were successfully annotated using SwissProt, Pfam, GO, and KEGG databases. A search against APD3, CAMP and LAMP databases identified 63 AMP candidates belonging to more than 20 diverse families and functional classes. mRNA of AvBD-2, AvBD-13 and CATH-2 were found to be differentially expressed between the three tested crows as well as among the tissues. We also characterized Corvus Cathelicidin 2 (CATH-2) to gain knowledge of its antimicrobial mechanisms. The CD spectroscopy of synthesized mature Corvus CATH-2 peptide displayed an amphipathic α-helical structure. Though the synthetic CATH-2 caused hemolysis of human RBC, it also exhibited antimicrobial activity against E. coli, S. aureus, and B. cereus. Docking simulation results revealed that this peptide could bind to the LPS binding site of MD-2, which may prevent LPS from entering the MD-2 binding pocket, and trigger TLR4 signaling pathway. The Corvus CATH-2 characterized in this study could aid in the development of novel therapeutics.
多药耐药性已成为与高发病率和死亡率相关的全球健康问题。抗菌肽已被认为是有前途的抗感染药物的潜在先导化合物。由于它们的清道夫生活方式，红嘴山鸦被认为对其饮食中的病原体产生了强大的免疫力，这意味着它们已经进化出了抵抗感染的机制。在本研究中，对红嘴山鸦的转录组进行了测序，并从头组装以鉴定抗菌肽基因的存在。获得了 7209 万条高质量的清洁读数，然后将其从头组装成 343503 条转录本和 74958 条基因。约 37559 条基因成功地使用 SwissProt、Pfam、GO 和 KEGG 数据库进行了注释。对 APD3、CAMP 和 LAMP 数据库的搜索鉴定出 63 种 AMP 候选物，它们属于 20 多种不同的家族和功能类别。AvBD-2、AvBD-13 和 CATH-2 的 mRNA 在三种测试的乌鸦以及组织之间存在差异表达。我们还对 Corvus 素 2 (CATH-2) 进行了表征，以了解其抗菌机制。合成的成熟 Corvus CATH-2 肽的 CD 光谱显示出两亲性 α-螺旋结构。尽管合成的 CATH-2 导致人 RBC 溶血，但它也对大肠杆菌、金黄色葡萄球菌和蜡样芽孢杆菌表现出抗菌活性。对接模拟结果表明，该肽可与 MD-2 的 LPS 结合位点结合，从而阻止 LPS 进入 MD-2 结合口袋，并触发 TLR4 信号通路。本研究中表征的 Corvus CATH-2 可有助于开发新型治疗药物。
