Flavonoid-mediated inhibition of SARS coronavirus 3C-like protease expressed in Pichia pastoris
Flavonoid-mediated inhibition of SARS coronavirus 3C-like protease expressed in Pichia pastoris
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DOI：
10.1007/s10529-011-0845-8
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发表时间：
2012-05-01
影响因子：
2.7
通讯作者：
Kim, Doman
中科院分区：
文献类型：
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作者：
Thi Thanh Hanh Nguyen;Woo, Hye-Jin;Kim, Doman
严重急性呼吸综合征相关冠状病毒（SARS-CoV）的3C样蛋白酶（3CL（pro））在SARS-CoV复制过程中起重要作用，是一个很有前途的药物靶点。重组3CL（pro）在GS 115中表达为42 kDa蛋白，其显示15 +/-Δ 2 μ M的α，以Dabcyl-KTSAVLQSGFRKME-Edans作为底物。纯化的3CL（pro）用于与七种黄酮类化合物的抑制和动力学测定。6种黄酮类化合物的IC_（50）为47-381 μ M。槲皮素、表没食子儿茶素没食子酸酯和没食子儿茶素没食子酸酯（GCG）对3CL（pro）表现出良好的抑制作用，IC 50值分别为73、73和47 μ M。GCG表现出竞争性抑制模式，其值为25 +/- A 1.7 μ M。在分子对接实验中，GCG与3CL（pro）活性位点的结合能为-14kcalmol（-1），其3-OH位的没食子酰基部分是3CL（pro）抑制活性所必需的。
The 3C-like protease (3CL(pro)) of severe acute respiratory syndrome associated coronavirus (SARS-CoV) is vital for SARS-CoV replication and is a promising drug target. Recombinant 3CL(pro) was expressed in GS115 as a 42 kDa protein that displayed a of 15 +/- A 2 mu M with Dabcyl-KTSAVLQSGFRKME-Edans as substrate. Purified 3CL(pro) was used for inhibition and kinetic assays with seven flavonoid compounds. The IC50 of six flavonoid compounds were 47-381 mu M. Quercetin, epigallocatechin gallate and gallocatechin gallate (GCG) displayed good inhibition toward 3CL(pro) with IC50 values of 73, 73 and 47 mu M, respectively. GCG showed a competitive inhibition pattern with value of 25 +/- A 1.7 mu M. In molecular docking experiments, GCG displayed a binding energy of -14 kcal mol(-1) to the active site of 3CL(pro) and the galloyl moiety at 3-OH position was required for 3CL(pro) inhibition activity.