Table_1_Proteomic Mapping of Multifunctional Complexes Within Triatomine Saliva.XLSX

Triatomines are hematophagous insects that transmit Trypanosoma cruzi, the etiological agent of Chagas disease. This neglected tropical disease represents a global health issue as it is spreading worldwide. The saliva of Triatominae contains miscellaneous proteins crucial for blood feeding acquisition, counteracting host's hemostasis while performing vasodilatory, anti-platelet and anti-coagulant activities, besides modulating inflammation and immune responses. Since a set of biological processes are mediated by protein complexes, here, the sialocomplexomes (salivary protein complexes) of five species of Triatominae were studied to explore the protein-protein interaction networks. Salivary multiprotein complexes from Triatoma infestans, Triatoma dimidiata, Dipetalogaster maxima, Rhodnius prolixus, and Rhodnius neglectus were investigated by Blue-Native- polyacrylamide gel electrophoresis coupled with liquid chromatography tandem mass spectrometry. More than 70 protein groups, uncovering the landscape of the Triatominae salivary interactome, were revealed. Triabin, actin, thioredoxin peroxidase and an uncharacterized protein were identified in sialocomplexes of the five species, while hexamerin, heat shock protein and histone were identified in sialocomplexes of four species. Salivary proteins related to triatomine immunity as well as those required during blood feeding process such as apyrases, antigen 5, procalins, and nitrophorins compose different complexes. Furthermore, unique proteins for each triatomine species were revealed. This study represents the first Triatominae sialocomplexome reference to date and shows that the approach used is a reliable tool for the analysis of Triatominae salivary proteins assembled into complexes.

锥蝽亚科（Triatominae）昆虫为吸血性昆虫，可传播恰加斯病（Chagas disease）的病原体克氏锥虫（Trypanosoma cruzi）。该被忽视的热带病在全球范围内蔓延，已成为全球性公共卫生问题。锥蝽唾液中含有多种对血液摄取至关重要的蛋白质，这些蛋白可通过发挥血管舒张、抗血小板及抗凝血活性对抗宿主止血系统，同时调节炎症与免疫应答。由于诸多生物学过程由蛋白质复合体介导，本研究针对5种锥蝽亚科昆虫的唾液蛋白复合体组（sialocomplexomes，salivary protein complexes）展开研究，以解析其蛋白质相互作用网络。本研究采用蓝色天然聚丙烯酰胺凝胶电泳（Blue-Native-polyacrylamide gel electrophoresis）结合液相色谱-串联质谱（liquid chromatography tandem mass spectrometry）技术，对红带锥蝽（Triatoma infestans）、中分锥蝽（Triatoma dimidiata）、大锥蝽（Dipetalogaster maxima）、长红猎蝽（Rhodnius prolixus）及贫毛红猎蝽（Rhodnius neglectus）的唾液多蛋白复合体进行了分析。研究共鉴定出70余个蛋白质组群，揭示了锥蝽唾液相互作用组的全貌。在5个物种的唾液蛋白复合体中均鉴定出锥蝽素（Triabin）、肌动蛋白（actin）、硫氧还蛋白过氧化物酶（thioredoxin peroxidase）及1种未表征蛋白；在4个物种的唾液蛋白复合体中鉴定出六聚体蛋白（hexamerin）、热休克蛋白（heat shock protein）及组蛋白（histone）。与锥蝽免疫及血液摄取过程相关的唾液蛋白，如腺苷三磷酸双磷酸酶（apyrases）、抗原5（antigen 5）、前钙蛋白（procalins）及硝合蛋白（nitrophorins），共同构成了不同的蛋白复合体。此外，本研究还发现了各锥蝽物种特有的蛋白质。本研究为目前首个锥蝽亚科唾液蛋白复合体组的参考数据集，证实所采用的技术方法可作为分析锥蝽唾液组装复合体蛋白的可靠工具。