中国毛虾流感病毒神经氨酸酶抑制活性肽的研究  

Active Peptide from Acetes Chinensis with Inhibitory Activity on Neuraminidase of Influenza Virus

作　　者：王海涛[1] 王伟 陈铭[1] 苑宁[1] 赵元晖[1] 毛相朝[1] 

机构地区：[1]中国海洋大学食品科学与工程学院 [2]医药学院,青岛266003

出　　处：《高等学校化学学报》2013年第11期2540-2545,共6页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:31000830);中央高校基本科研业务费专项资金(批准号:201262021);长江学者和创新团队发展计划(批准号:IRT1188)资助

摘　　要：以中国毛虾为原料,以抑制流感病毒神经氨酸酶(NA)活性为初筛指标,通过控制酶切位点制备了具有抑制NA活性的酶解液.利用凝胶层析和高效液相色谱等技术分离纯化出高活性的抑制肽,其IC50值为96.1μmol/L.经串联质谱测定该抑制肽序列为EISYIHAEAYRRGELK,紫外光谱分析结果证明该抑制肽能与NA结合.基于反向对接,应用SYBYL软件模拟抑制肽与NA活性区域结合,确定了抑制肽与NA的结合位点.细胞毒性实验测得该抑制肽对细胞的最大无毒浓度(TC0)为1.26 mg/mL.在红细胞凝集实验中,随着抑制肽浓度增大,病毒的凝集价显著降低,证明抑制肽的抗病毒作用具有多靶点.Low-valued Acetes Chinensis was chosen to prepare the enzymatic hydrolysate with neuraminidase(NA) inhibitory activity by controlled hydrolysis. A novel NA inhibitory peptide was purified by gel chromatography and reversed phase high performance liquid chromatography. The IC50value of the peptide was only96. 1 μmol / L and it could also significantly inhibit influenza A virus replication in MDCK cells. The amino acid sequence of this peptide was EISYIHAEAYRRGELK which identified by LC-MS / MS and its binding with NA protein was proved by UV spectrum analysis. The accurate sites of NA protein that the peptide bind to were determined using the computer assisted software SYBYL. The largest non-toxic concentration( TC0)was1. 26 mg / mL which assayed by cell cytotoxicity experiments. The hemagglutination essay showed that this pepetide could obviously decrease the agglutination titer influenza virus in a dose-dependent manner which suggested that the antiviral activity of this peptide might be attributed to its action on multiple targets.

关 键 词：中国毛虾 神经氨酸酶 NA抑制肽 分离纯化 NA结合位点 

分 类 号：O629.72[理学—有机化学]