Division of Microbiology and Immunology, Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Department of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan.
J Biol Chem. 2022 Jun;298(6):101957. doi: 10.1016/j.jbc.2022.101957. Epub 2022 Apr 20.
Japanese encephalitis is a mosquito-borne disease caused by the Japanese encephalitis virus (JEV) that is prevalent in Asia and the Western Pacific. Currently, there is no effective treatment for Japanese encephalitis. Curcumin (Cur) is a compound extracted from the roots of Curcuma longa, and many studies have reported its antiviral and anti-inflammatory activities. However, the high cytotoxicity and very low solubility of Cur limit its biomedical applications. In this study, Cur carbon quantum dots (Cur-CQDs) were synthesized by mild pyrolysis-induced polymerization and carbonization, leading to higher water solubility and lower cytotoxicity, as well as superior antiviral activity against JEV infection. We found that Cur-CQDs effectively bound to the E protein of JEV, preventing viral entry into the host cells. In addition, after continued treatment of JEV with Cur-CQDs, a mutant strain of JEV was evolved that did not support binding of Cur-CQDs to the JEV envelope. Using transmission electron microscopy, biolayer interferometry, and molecular docking analysis, we revealed that the S123R and K312R mutations in the E protein play a key role in binding Cur-CQDs. The S123 and K312 residues are located in structural domains II and III of the E protein, respectively, and are responsible for binding to receptors on and fusing with the cell membrane. Taken together, our results suggest that the E protein of flaviviruses represents a potential target for the development of CQD-based inhibitors to prevent or treat viral infections.
日本脑炎是一种由乙型脑炎病毒(JEV)引起的蚊媒疾病,流行于亚洲和西太平洋地区。目前,尚无有效的日本脑炎治疗方法。姜黄素(Cur)是从姜黄属植物的根部提取的一种化合物,许多研究报道了其抗病毒和抗炎活性。然而,Cur 的高细胞毒性和极低的溶解度限制了其在生物医学中的应用。在本研究中,通过温和的热解诱导聚合和碳化合成了姜黄素碳量子点(Cur-CQDs),从而提高了水溶解度和降低了细胞毒性,并具有针对 JEV 感染的优越抗病毒活性。我们发现 Cur-CQDs 可有效地与 JEV 的 E 蛋白结合,阻止病毒进入宿主细胞。此外,在继续用 Cur-CQDs 处理 JEV 后,进化出了一种不支持 Cur-CQDs 与 JEV 包膜结合的 JEV 突变株。通过透射电子显微镜、生物层干涉测量法和分子对接分析,我们揭示了 E 蛋白中的 S123R 和 K312R 突变在结合 Cur-CQDs 方面起着关键作用。S123 和 K312 残基分别位于 E 蛋白的结构域 II 和 III 中,负责与细胞膜上的受体结合并融合。总之,我们的研究结果表明,黄病毒的 E 蛋白代表了开发基于 CQD 的抑制剂以预防或治疗病毒感染的潜在靶标。