Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea.
The Institute of Food Industrialization, Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do 25354, Republic of Korea.
J Microbiol Biotechnol. 2023 Jun 28;33(6):788-796. doi: 10.4014/jmb.2211.11040. Epub 2023 Feb 13.
Canine parvovirus type 2 (CPV-2) has high morbidity and mortality rates in canines. Nonstructural protein 1 (NS1) of CPV-2 has endonuclease activity, initiates viral DNA replication, and is highly conserved. Thus, it is a promising target for antiviral inhibitor development. We overexpressed a 41.9 kDa active recombinant endonuclease in and designed a nicking assay using carboxyfluorescein and quencher-linked ssDNA as substrates. The optimal temperature and pH of the endonuclease were 37°C and pH 7, respectively. Curcumin, bisdemethoxycurcumin, demethoxycurcumin, linoleic acid, tannic acid, and α-tocopherol inhibited CPV-2 NS1 endonuclease with IC values of 0.29 to 8.03 μM. The extracted turmeric, yerba mate, and sesame cake suppressed CPV-2 NS1 endonuclease with IC values of 1.48, 7.09, and 52.67 μg/ml, respectively. The binding affinity between curcumin, the strongest inhibitor, and CPV-2 NS1 endonuclease by molecular docking was -6.4 kcal/mol. Curcumin inhibited CPV-2 NS1 endonuclease via numerous hydrophobic interactions and two hydrogen bonds with Lys97 and Pro111 in the allosteric site. These results suggest that adding curcuminoids, linoleic acid, tannic acid, α-tocopherol, extracted turmeric, sesame cake, and yerba to the diet could prevent CPV-2 infection.
犬细小病毒 2 型（CPV-2）在犬中具有高发病率和死亡率。CPV-2 的非结构蛋白 1（NS1）具有内切酶活性，启动病毒 DNA 复制，且高度保守。因此，它是开发抗病毒抑制剂的有前途的靶标。我们在 中过表达了具有 41.9 kDa 活性的重组内切酶，并设计了一种使用羧基荧光素和猝灭剂连接的 ssDNA 作为底物的切口测定法。内切酶的最佳温度和 pH 值分别为 37°C 和 pH 7。姜黄素、双去甲氧基姜黄素、去甲氧基姜黄素、亚油酸、单宁酸和 α-生育酚对 CPV-2 NS1 内切酶的抑制作用的 IC 值为 0.29 至 8.03 μM。从姜黄、马黛茶和芝麻饼中提取的物质对 CPV-2 NS1 内切酶的抑制作用的 IC 值分别为 1.48、7.09 和 52.67 μg/ml。通过分子对接，姜黄素与 CPV-2 NS1 内切酶之间的结合亲和力为-6.4 kcal/mol。姜黄素通过与别构位点中的 Lys97 和 Pro111 形成两个氢键和许多疏水相互作用来抑制 CPV-2 NS1 内切酶。这些结果表明，在饮食中添加姜黄素类、亚油酸、单宁酸、α-生育酚、提取的姜黄、芝麻饼和马黛茶可能会预防 CPV-2 感染。
