Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
J Proteomics. 2022 Mar 15;254:104476. doi: 10.1016/j.jprot.2021.104476. Epub 2022 Jan 4.
Over the last 20 years, advances in sequencing technologies paired with biochemical and structural studies have shed light on the unique pharmacological arsenal produced by the salivary glands of hematophagous arthropods that can target host hemostasis and immune response, favoring blood acquisition and, in several cases, enhancing pathogen transmission. Here we provide a deeper insight into Xenopsylla cheopis salivary gland contents pairing transcriptomic and proteomic approaches. Sequencing of 99 pairs of salivary glands from adult female X. cheopis yielded a total of 7432 coding sequences functionally classified into 25 classes, of which the secreted protein class was the largest. The translated transcripts also served as a reference database for the proteomic study, which identified peptides from 610 different proteins. Both approaches revealed that the acid phosphatase family is the most abundant salivary protein group from X. cheopis. Additionally, we report here novel sequences similar to the FS-H family, apyrases, odorant and hormone-binding proteins, antigen 5-like proteins, adenosine deaminases, peptidase inhibitors from different subfamilies, proteins rich in Glu, Gly, and Pro residues, and several potential secreted proteins with unknown function. SIGNIFICANCE: The rat flea X. cheopis is the main vector of Yersinia pestis, the etiological agent of the bubonic plague responsible for three major pandemics that marked human history and remains a burden to human health. In addition to Y. pestis fleas can also transmit other medically relevant pathogens including Rickettsia spp. and Bartonella spp. The studies of salivary proteins from other hematophagous vectors highlighted the importance of such molecules for blood acquisition and pathogen transmission. However, despite the historical and clinical importance of X. cheopis little is known regarding their salivary gland contents and potential activities. Here we provide a comprehensive analysis of X. cheopis salivary composition using next generation sequencing methods paired with LC-MS/MS analysis, revealing its unique composition compared to the sialomes of other blood-feeding arthropods, and highlighting the different pathways taken during the evolution of salivary gland concoctions. In the absence of the X. cheopis genome sequence, this work serves as an extended reference for the identification of potential pharmacological proteins and peptides present in flea saliva.
在过去的 20 年中，测序技术的进步与生化和结构研究相结合，揭示了吸血节肢动物唾液腺产生的独特药理学武器库，这些武器库可以靶向宿主止血和免疫反应，有利于血液获取，并且在某些情况下，增强病原体的传播。在这里，我们通过转录组学和蛋白质组学方法深入了解了印鼠客蚤唾液腺的内容物。对 99 对成年雌性印鼠客蚤唾液腺的测序总共产生了 7432 个编码序列，这些序列根据功能分为 25 类，其中分泌蛋白类最大。翻译后的转录本也作为蛋白质组学研究的参考数据库，该研究鉴定了来自 610 种不同蛋白质的肽。这两种方法都表明，酸性磷酸酶家族是印鼠客蚤中最丰富的唾液蛋白组。此外，我们在这里报告了类似于 FS-H 家族、apyrases、气味和激素结合蛋白、抗原 5 样蛋白、腺苷脱氨酶、不同亚家族的肽酶抑制剂、富含 Glu、Gly 和 Pro 残基的蛋白质以及几种具有未知功能的潜在分泌蛋白的新序列。意义：印鼠客蚤是鼠疫耶尔森菌的主要媒介，鼠疫耶尔森菌是引起腺鼠疫的病原体，腺鼠疫曾导致三次大流行，标志着人类历史，并仍然对人类健康构成负担。除鼠疫耶尔森菌外，跳蚤还可以传播其他与医学相关的病原体，包括立克次体和巴尔通体。其他吸血节肢动物唾液蛋白的研究强调了这些分子对血液获取和病原体传播的重要性。然而，尽管印鼠客蚤具有历史和临床重要性，但人们对其唾液腺内容物和潜在活动知之甚少。在这里，我们使用下一代测序方法与 LC-MS/MS 分析相结合，对印鼠客蚤唾液成分进行了全面分析，与其他吸血节肢动物的唾液组相比，揭示了其独特的组成，并强调了在唾液腺混合物的进化过程中所采取的不同途径。在没有印鼠客蚤基因组序列的情况下，这项工作为鉴定跳蚤唾液中存在的潜在药理蛋白和肽提供了扩展参考。
