Research Center for Parasites & Vectors (RCPV), College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China.
Research Center for Parasites & Vectors (RCPV), College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China.
Vet Parasitol. 2023 Jun;318:109933. doi: 10.1016/j.vetpar.2023.109933. Epub 2023 Apr 7.
Tick saliva is a reservoir of bioactive proteins. Saliva protein compositions change dynamically during blood-feeding. Decipherment of protein profiles in different blood-feeding stages may bring deeper insight into tick feeding physiology and provide targets for immunologic control alternatives. However, having the infancy of tick genome sequencing, assembly, annotation, and limited knowledge of tick salivary proteins restrain the data interpretation. Here, we aimed to depict the saliva protein profile in partially- (PE) and fully-engorged (FE) Haemaphysalis flava ticks, with a special focus on the analysis of those uncharacterized proteins. Saliva was collected from PE and FE adult female H. flava ticks. Saliva proteins were analyzed by high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS-MS). MS data were searched against an in-house salivary gland transcriptome library for identification of tick-derived proteins. Abundances of proteins were compared between PE and FE ticks. The uncharacterized proteins detected in saliva were further bioinformatically analyzed. In total, 614 proteins were identified including 94 host proteins and 520 tick-derived proteins. The 226 tick-derived high-confidence proteins were classified into 10 categories: transporters, enzymes, protease inhibitors, immunity-related proteins, lipocalins, glycine-rich proteins, muscle proteins, secreted proteins, uncharacterized proteins and others. A total of 98 proteins were shared in both PE and FE with 74 only in PE and 54 only in FE. Abundances of 24 shared proteins were significantly higher in PE. The profile of top 15 most abundant proteins was also different between PE and FE ticks. The 65 uncharacterized proteins detected in tick saliva were branched into subclusters 1 A, 1B, 2, 3 A, 3B and 3 C based on particular motifs like RGD, LRR, indicating their diverse predicted functions like anti-coagulation, regulation of innate immune, or other functions. This study provides and compares saliva proteomes of H. flava ticks in two feeding stages with special cluster analysis on the uncharacterized proteins. Further investigations are needed to confirm the roles of these uncharacterized proteins in ticks.
蜱唾液是生物活性蛋白的储库。在吸血过程中，唾液蛋白的组成会发生动态变化。解析不同吸血阶段的蛋白质谱可能会深入了解蜱的进食生理学，并为免疫控制替代物提供靶标。然而，由于蜱基因组测序、组装、注释的起步阶段，以及对蜱唾液蛋白的有限了解，限制了数据的解释。在这里，我们旨在描述部分（PE）和完全饱血（FE）的中华硬蜱的唾液蛋白谱，特别关注那些未被描述的蛋白质的分析。从 PE 和 FE 成年雌性中华硬蜱中收集唾液。通过高效液相色谱-串联质谱（HPLC/MS-MS）分析唾液蛋白。将 MS 数据与内部唾液腺转录组文库进行比对，以鉴定源自蜱的蛋白质。比较 PE 和 FE 蜱之间蛋白质的丰度。对唾液中检测到的未被描述的蛋白质进行进一步的生物信息学分析。总共鉴定出 614 种蛋白质，包括 94 种宿主蛋白和 520 种源自蜱的蛋白质。226 种源自蜱的高可信度蛋白质被分为 10 类：转运蛋白、酶、蛋白酶抑制剂、免疫相关蛋白、脂联素、富含甘氨酸的蛋白、肌肉蛋白、分泌蛋白、未被描述的蛋白和其他。PE 和 FE 中共有 98 种蛋白，PE 中仅 74 种，FE 中仅 54 种。24 种共享蛋白的丰度在 PE 中显著更高。PE 和 FE 蜱之间的前 15 种最丰富蛋白的图谱也不同。在蜱唾液中检测到的 65 种未被描述的蛋白根据特定的基序（如 RGD、LRR）分为亚簇 1A、1B、2、3A、3B 和 3C，表明它们具有多样化的预测功能，如抗凝血、调节先天免疫或其他功能。本研究提供并比较了两种进食阶段的中华硬蜱的唾液蛋白质组，并对未被描述的蛋白质进行了特殊的聚类分析。需要进一步的研究来确认这些未被描述的蛋白质在蜱中的作用。
