Key Laboratory of Livestock Infectious Diseases, Ministry of Education, and Key Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, 120 Dongling Road, Shenyang, 110866, China.
Research Unit for Pathogenic Mechanisms of Zoonotic Parasites, Chinese Academy of Medical Sciences, 120 Dongling Road, Shenyang, 110866, China.
Parasit Vectors. 2024 Mar 4;17(1):105. doi: 10.1186/s13071-024-06207-7.
The human sortilin protein is an important drug target and detection marker for cancer research. The sortilin from Toxoplasma gondii transports proteins associated with the apical organelles of the parasite. In this study, we aimed to determine the intracellular localization and structural domains of T. gondii sortilin, which may mediate protein transportation. Approaches to the functional inhibition of sortilin to establish novel treatments for T. gondii infections were explored.
A gene encoding the sortilin protein was identified in the T. gondii genome. Immunoprecipitation and mass spectrometry were performed to identify the protein species transported by T. gondii sortilin. The interaction of each structural domain of sortilin with the transported proteins was investigated using bio-layer interferometry. The binding regions of the transported proteins in sortilin were identified. The effect of the sortilin inhibitor AF38469 on the infectivity of T. gondii was investigated. The binding site of AF38469 on sortilin was determined.
The subdomains Vps10, sortilin-C, and sortilin-M of the sortilin were identified as the binding regions for intracellular transportation of the target proteins. The sortilin inhibitor AF38469 bound to the Vps10 structural domain of T. gondii sortilin, which inhibited parasite invasion, replication, and intracellular growth in vitro and was therapeutic in mice infected with T. gondii.
The Vps10, sortilin-C, and sortilin-M subdomains of T. gondii sortilin were identified as functional regions for intracellular protein transport. The binding region for the sortilin inhibitor AF38469 was also identified as the Vps10 subdomain. This study establishes sortilin as a promising drug target against T. gondii and provides a valuable reference for the development of anti-T. gondii drug-target studies.
人源分选连接蛋白（Sortilin）是癌症研究的重要药物靶点和检测标志物。刚地弓形虫分选连接蛋白（TgSortilin）可转运与寄生虫顶器相关的蛋白。本研究旨在确定 TgSortilin 的细胞内定位和结构域，这些结构域可能介导蛋白转运。我们还探索了通过分选连接蛋白的功能抑制来建立弓形虫感染新疗法的方法。
在弓形虫基因组中鉴定出分选连接蛋白的基因。通过免疫沉淀和质谱分析鉴定 TgSortilin 转运的蛋白种类。使用生物层干涉法研究分选连接蛋白各结构域与转运蛋白的相互作用。鉴定分选连接蛋白中转运蛋白的结合区。研究分选连接蛋白抑制剂 AF38469 对弓形虫感染性的影响。确定 AF38469 在分选连接蛋白上的结合位点。
鉴定出分选连接蛋白的 Vps10、sortilin-C 和 sortilin-M 亚结构域为靶蛋白细胞内转运的结合区。分选连接蛋白抑制剂 AF38469 与弓形虫分选连接蛋白的 Vps10 结构域结合，抑制寄生虫体外侵袭、复制和细胞内生长，并在感染弓形虫的小鼠中具有治疗作用。
确定了弓形虫分选连接蛋白的 Vps10、sortilin-C 和 sortilin-M 亚结构域为细胞内蛋白转运的功能区域。还确定了分选连接蛋白抑制剂 AF38469 的结合区为 Vps10 亚结构域。本研究确立了分选连接蛋白作为抗弓形虫药物靶点的潜力，并为抗弓形虫药物靶点研究的发展提供了有价值的参考。
