囊状幼虫病病毒衣壳蛋白肝素结合基序鉴定及其抗感染作用分析  

Identification and anti-infection effect of heparin binding motif of sacbrood virus capsid protein

作　　者：沈克飞[1] 曹兰[1] 徐登峰[1] 许国洋 杨金龙[1] 罗文华[1] SHEN Kefei;CAO Lan;XU Dengfeng;XU Guoyang;YANG Jinlong;LUO Wenhua(Chongqing Academy of Animal Sciences,Chongqing 402460,China)

机构地区：[1]重庆市畜牧科学院,重庆402460

出　　处：《中国生物制品学杂志》2023年第12期1465-1470,1475,共7页Chinese Journal of Biologicals

基　　金：重庆市自然科学基金面上项目(cstc2020jcyj-msxmX1047);重庆市现代农业产业技术体系创新团队(蜂业)(CQMAITS202316)。

摘　　要：目的鉴定囊状幼虫病病毒(sacbrood virus,SBV)药物结合口袋蛋白2(drug-binding pocket protein 2,Dbpp2)的肝素结合基序,并测定肝素结合基序及其衍生肽的抗SBV感染活性。方法采用在线软件对Dbpp2进行二级结构预测和同源建模,以搜索肝素结合基序,利用肝素琼脂糖珠检测SBV、Dbpp2及其肝素结合基序突变体与肝素的结合能力、肝素结合基序抑制Dbpp2结合肝素的能力,并检测肝素结合基序及其衍生肽对SBV感染的中华蜜蜂(Apis cerana cerana)幼虫的治疗效果。结果SBV和Dbpp2可结合肝素,Dbpp2 C-端暴露在表面的富含碱性氨基酸的柔性环(KPANRPRR)为肝素结合基序,该基序可抑制Dbpp2结合肝素。KPANRPRR及其衍生肽KPAARPRR、KPRN-RPRR、KPRARPRR、KPRWRPRR、KPRWRPRRW均具有降低SBV感染导致幼虫死亡率的作用,其中,KPRARPRR的治疗效果最佳。结论本研究为阐明SBV-氨基萄聚糖(glycosaminoglycan,GAG)相互作用奠定了基础,也为基于相互作用开发治疗SBV感染的多肽提供了参考。Objective To identify the heparin-binding motif of drug-binding pocket protein 2(Dbpp2)of sacbrood virus(SBV)and determine the anti-SBV infection activity of the heparin-binding motif and its derived peptides.Methods The secondary structure prediction and homology modeling of Dbpp2 were carried out using online software to search for potential heparin-binding motifs.The binding ability of SBV,Dbpp2 and their heparin-binding motif mutants to and the ability of heparin-binding motifs to inhibit the binding of Dbpp2 to heparin were tested using heparin agarose beads.The therapeutic effect of heparin-binding motifs and their derived peptides on the larvae of Apis cerana cerana infected with SBV was tested.Results SBV and Dbpp2 showed binding with heparin.The flexible ring(KPANRPRR)rich in basic amino acids exposed at the C-end of Dbpp2 was a heparin-binding motif,which inhibited Dbpp2 from binding to heparin.KPANRPRR and its derived peptides KPAARPRR,KPRNRPRR,KPRARPRR,KPRWRPRR and KPRWRPRRW all had the effect of reducing the mortality of larvae caused by SBV infection,among which KPRARPRR showed the best therapeutic effect.Conclusion The research provides a basis for understanding the interaction of SBV-glycosaminoglycan(GAG),and provides a reference for the development of peptides for the treatment of SBV infection based on the interaction.

关 键 词：囊状幼虫病病毒 衣壳蛋白 药物结合口袋 肝素结合基序 

分 类 号：S895.133[农业科学—特种经济动物饲养]