Department of Public Health and Infectious Diseases, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy.
National Center for Innovative Technologies in Public Health, Italian National Institute of Health, Viale Regina Elena 299, 00161 Rome, Italy.
Int J Mol Sci. 2024 May 18;25(10):5529. doi: 10.3390/ijms25105529.
Mosquito saliva plays a crucial physiological role in both sugar and blood feeding by helping sugar digestion and exerting antihemostatic functions. During meal acquisition, mosquitoes are exposed to the internalization of external microbes. Since mosquitoes reingest significant amounts of saliva during feeding, we hypothesized that salivary antimicrobial components may participate in the protection of mouthparts, the crop, and the gut by inhibiting bacterial growth. To identify novel potential antimicrobials from mosquito saliva, we selected 11 candidates from salivary transcriptomic datasets and obtained them either using a cell-free transcription/translation expression system or, when feasible, via chemical synthesis. Hyp6.2 and hyp13, which were predicted to be produced as propeptides and cleaved in shorter mature forms, showed the most interesting results in bacterial growth inhibition assays. Hyp6.2 (putative mature form, 35 amino acid residues) significantly inhibited the growth of Gram-positive () and Gram-negative ( and ) bacteria. Hyp13 (short form, 19 amino acid residues) dose-dependently inhibited and growth, inducing membrane disruption in both Gram-positive and Gram-negative bacteria as indicated with scanning electron microscopy. In conclusion, we identified two salivary peptides inhibiting Gram-positive and Gram-negative bacteria growth and possibly contributing to the protection of mosquito mouthparts and digestive tracts from microbial infection during and/or after feeding.
蚊子唾液在吸食糖和血液过程中发挥着至关重要的生理作用,有助于消化糖分并发挥抗凝血功能。在取食过程中,蚊子会接触到内部微生物的内化。由于蚊子在进食过程中会重新摄入大量唾液,我们假设唾液中的抗菌成分可能通过抑制细菌生长来保护口器、嗉囊和肠道。为了从蚊子唾液中鉴定出新型潜在的抗菌物质,我们从唾液转录组数据集选择了 11 个候选物,并使用无细胞转录/翻译表达系统获得它们,或者在可行的情况下通过化学合成获得。Hyp6.2 和 Hyp13 被预测为前肽并以较短的成熟形式切割,在细菌生长抑制试验中显示出最有趣的结果。Hyp6.2(假定的成熟形式,35 个氨基酸残基)显著抑制革兰氏阳性菌(和)和革兰氏阴性菌(和)的生长。Hyp13(短形式,19 个氨基酸残基)以剂量依赖的方式抑制 和 生长,扫描电子显微镜显示其在革兰氏阳性菌和革兰氏阴性菌中均诱导了膜破裂。总之,我们鉴定出两种 唾液肽可抑制革兰氏阳性菌和革兰氏阴性菌的生长,并可能有助于保护蚊子口器和消化道免受微生物感染,无论是在取食期间还是取食后。