Phage display library based antiviral screening:porcine respiratory and reproductive syndrome virus endoribonuclease is a target for antiviral screening
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- 英文论文题名：Phage display library based antiviral screening:porcine respiratory and reproductive syndrome virus endoribonuclease is a target for antiviral screening
- 论文作者：Ke LIU ; Donghua SHAO ; Beili HUAN ; Beibei LI ; Yafeng QIU ; Yuanyuan SHI ; Jianchao WEI ; Zhiyong MA
- 英文论文作者：Ke LIU ; Donghua SHAO ; Beili HUAN ; Beibei LI ; Yafeng QIU ; Yuanyuan SHI ; Jianchao WEI ; Zhiyong MA ; Shanghai Veterinary Research Institute ; Chinese Academy of Agricultural Science ; Key Laboratory of Animal Disease Diagnostic & Immunology ; Department of Veterinary Medicine College ; Nanjing Agricultural University ; Shanghai Academy of Agricultural Sciences
- 年：2016
- 作者机构：Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Science;Key Laboratory of Animal Disease Diagnostic & Immunology,Department of Veterinary Medicine College,Nanjing Agricultural University;Shanghai Academy of Agricultural Sciences;
- 英文论文关键词：Viral endoribonuclease ; phage display library ; porcine reproductive and respiratory syndrome virus ; antiviral screening
- 会议召开时间：2016-07-20
- 会议录名称：中国畜牧兽医学会兽医公共卫生学分会第五次学术研讨会论文集
- 语种：英文
- 分类号：S852.651
- 学会代码：SYGW
- 会议名称：中国畜牧兽医学会兽医公共卫生学分会第五次学术研讨会
- 会议地点：中国内蒙古通辽
- 主办单位：中国畜牧兽医学会兽医公共卫生学分会
- 学会名称：中国畜牧兽医学会兽医公共卫生学分会
- 页数：14
- 文件大小：849k
- 原文格式：D
- 会议级别：全国
摘要
Viral endoribonuclease plays a vital role in the physiological and biochemical activities of viral RNA.In our studies on PRRSV,we seek to screen candidate antiviral molecules that target this endoribonuclease.Through5 rounds of phage-display screening of 12-ammino acid peptides,4 peptides that show high affinity to the viral endoribonuclease were identified.In vitro experiments showed very high antiviral activity of all these 4 peptides.Wherein,the E4 peptide(NIPIKPRPRLMK) showed the highest antiviral activity(IC50 = 39.21μM),yet the lowest cytotoxicity.E4 can directly bind to the endoribonuclease of PRRSV(NSP11) and reduce copy number of viral RNA by 23.68 folds.Through large-scale phage-display screening and following in vitro analysis,we proved that viral endoribonuclease is a potential target of antiviral drugs.
Viral endoribonuclease plays a vital role in the physiological and biochemical activities of viral RNA.In our studies on PRRSV,we seek to screen candidate antiviral molecules that target this endoribonuclease.Through5 rounds of phage-display screening of 12-ammino acid peptides,4 peptides that show high affinity to the viral endoribonuclease were identified.In vitro experiments showed very high antiviral activity of all these 4 peptides.Wherein,the E4 peptide(NIPIKPRPRLMK) showed the highest antiviral activity(IC50 = 39.21μM),yet the lowest cytotoxicity.E4 can directly bind to the endoribonuclease of PRRSV(NSP11) and reduce copy number of viral RNA by 23.68 folds.Through large-scale phage-display screening and following in vitro analysis,we proved that viral endoribonuclease is a potential target of antiviral drugs.
Viral endoribonuclease plays a vital role in the physiological and biochemical activities of viral RNA.In our studies on PRRSV,we seek to screen candidate antiviral molecules that target this endoribonuclease.Through5 rounds of phage-display screening of 12-ammino acid peptides,4 peptides that show high affinity to the viral endoribonuclease were identified.In vitro experiments showed very high antiviral activity of all these 4 peptides.Wherein,the E4 peptide(NIPIKPRPRLMK) showed the highest antiviral activity(IC50 = 39.21μM),yet the lowest cytotoxicity.E4 can directly bind to the endoribonuclease of PRRSV(NSP11) and reduce copy number of viral RNA by 23.68 folds.Through large-scale phage-display screening and following in vitro analysis,we proved that viral endoribonuclease is a potential target of antiviral drugs.
引文
[1]Meulenberg,J.J.,Hulst,M.M.,Meijer,D.E.,Moonen,E.J.,Den,B.P.L.,Kluyver,D.E.1993.Lelystad virus,the causative agent of porcine epidemic abortion and respiratory syndrome(PEARS),is related to LDV and EAV.Virology 192:62-72.
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[3]Meng,X.J.,Paul,P.S.,Halbur,P.G.,Lum,M.A.1995.Phylogenetic analyses of the putative M(ORF 6)and N(ORF 7)genes of porcine reproductive and respiratory syndrome virus(PRRSV):implication for the existence of two genotypes of PRRSV in the USA and Europe.Archives of Virology 140:745-55.
[4]Mounir,S.,Mardassi,H.,Dea,S.1995.Identification and characterization of the porcine reproductive and respiratory virus ORFs 7,5 and 4 products.Advances in Experimental Medicine and Biology 380:317-20.
[5]Nelson,E.A.,Christopher-Hennings,J.,Benfield,D.A.1995.Structural proteins of porcine reproductive and respiratory syndrome virus(PRRSV).Advances in Experimental Medicine and Biology 380:321-23.
[6]Fang,Y.,Kim,D-Y.,Ropp,S.,Steen,P.,Christopher-Hennings,J.,Nelson,E.A.2004.Heterogeneity in Nsp2of European-like porcine reproductive and respiratory syndrome viruses isolated in the United States.Virus Research 100:229-35.
[7]Cafruny,W.A.,Duman,R.G.,Wong,G.H.W.,Said,S.,Ward-Demo,P.,Rowland,R.R.R.2006.Porcine reproductive and respiratory syndrome virus(PRRSV)infection spreads by cell-to-cell transfer in cultured MARC-145 cells,is dependent on an intact cytoskeleton,and is suppressed by drug-targeting of cell permissiveness to virus infection.Virology Journal 3:90.
[8]Bai,F.W.,Town,T.,Pradhan,D.,Cox,J.,Ashish,Ledizet,M.,Anderson,J.F.,Flavell,R.A.,Krueger,J.K.,Koski,R.A.2006.Antiviral Peptides Targeting the West Nile Virus Envelope Protein.Journal of Virology81:2047-55.
[9]Bajorath,J.,Peltason,L.,Wawer,M.,Guha,R.,Lajiness,M.S.,Van Drie,J.H.2009.Navigating structure-activity landscapes.Drug Discovery Today 14:698-705.
[10]Hall,P.R.,Hjelle,B.,Njus,H.,Ye,C.,Bondu-Hawkins,V.,Brown,D.C.2009.Phage Display Selection of Cyclic Peptides That Inhibit Andes Virus Infection.Journal of Virology 83:8965-69.
[11]Hancock,R.E.W.,Sahl,H-G.2006.Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies.Nature Biotechnology 24:1551-57.
[12]Ho,K.L.,Yusoff,K.,Seow,H.F.,Tan,W.S.,2003.Selection of high affinity ligands to hepatitis B core antigen from a phage-displayed cyclic peptide library.Journal of Medical Virology 69:27-32.
[13]Peng,B.,Chen,H.,Tan,Y.,Jin,M.,Guo,A.2006.Identification of one peptide which inhibited infectivity of avian infectious bronchitis virus in vitro.Science China Life Sciences 49:158-163.
[14]Sergeeva,A.,Kolonin,M.,Molldrem,J.,Pasqualini,R.,Arap,W.2006.Display technologies:Application for the discovery of drug and gene delivery agents.Advanced Drug Delivery Reviews 58:1622-54.
[15]N.Zou,J.Xia,F.Wang,Z.Duan,D.Miao,Q.Yan,S.Cao,X.Wen,P.Liu,Y.Huang.2015.Two novel neutralizing antigenic epitopes of the s1 subunit protein of a QX-like avian infectious bronchitis virus strain Sczy3as revealed using a phage display peptide library.Veterinary immunology and immunopathology,published online Epub Aug 19.
[16]Y.Du,L.Hou,J.Li.2000.Epitope mapping of HIV-1 using phage-display random peptide library and the purified Ig G from HIV patient.Chinese journal of experimental and clinical virology 14:338-341.
[17]Z.Zhang,L.Xu,Z.Wang.2011.Screening serum biomarkers for early primary hepatocellular carcinoma using a phage display technique.Journal of clinical laboratory analysis 25:402-408.
[18]Q.Deng,M.Zhuang,Y.Y.Kong,Y.H.Xie,Y.Wang.2005.Screening for Pre S specific binding ligands with a phage displayed peptides library.World journal of gastroenterology 11:4018-4023.
[19]Z.Han,F.Zhao,Y.Shao,X.Liu,X.Kong,Y.Song,S.Liu.2013.Fine level epitope mapping and conservation analysis of two novel linear B-cell epitopes of the avian infectious bronchitis coronavirus nucleocapsid protein.Virus research 171:54-64.
[20]Y.W.Zhong,D.P.Xu,X.D.Li,J.Z.Dai,B.Xu,L.Li.2009.Epitope screening of influenza A(H3N2)by using phage display library,Chinese journal of experimental and clinical virology 23:272-274.
[21]H.Xu,J.Wang,Z.Yu,F.Lv,J.Hou.2011.Screening of polypeptides binding to porcine reproductive and respiratory syndrome virus by phage display library.Wei sheng wu xue bao=Acta microbiologica Sinica51:127-133
[22]Oleksiewicz,M.B.,Botner,A.,Madsen,K.G.,Storgaard,T.1998.Sensitive detection and typing of porcine reproductive and respiratory syndrome virus by RT-PCR amplification of whole viral genes.Veterinary Microbiology 64:7-22.
[23]Sachs L,A.,Schnurr,D.,Yagi,S.,Lachowicz-Scroggins,M.E.,Widdicombe,J.H.2011.Quantitative real-time PCR for rhinovirus,and its use in determining the relationship between TCID50 and the number of viral particles.Journal of Virological Methods 171:212-18.
[24]Shimozono,M.2008.Engineering FRET Constructs Using CFP and YFP.Methods in Cell Biology85:381-393.
[25]Shyu,J.Y.,Liu,H.,Deng,X.,Hu,C.D.2006.Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions.Biotechniques 40:61-66.
[26]Mateu,E.,Diaz,I.2008.The challenge of PRRS immunology.The Veterinary Journal 177:345-51.
[27]Storgaard,T.,Oleksiewicz,M.B.,Botner,A.1999.Examination of the selective pressures on a live PRRSvaccine virus.Archives of Virology 144:2389-2401.
[28]Y.Fang,E.J.Snijder.2010.The PRRSV replicase:exploring the multifunctionality of an intriguing set of nonstructural proteins.Virus research 154:61-76.
[29]Hagiwara,K.,Kondoh,Y.,Ueda,A.,Yamada,K.,Goto,H.,Watanabe,T.2010.Discovery of novel antiviral agents directed against the influenza A virus nucleoprotein using photo-cross-linked chemical arrays.Biochemical and Biophysical Research Communications 394:721-27.
[30]Y.Sun,D.Li,S.Giri,S.G.Prasanth,D.Yoo.2014.Differential host cell gene expression and regulation of cell cycle progression by nonstructural protein 11 of porcine reproductive and respiratory syndrome virus.Bio Med research international 2014:430508
[31]Hinson,E.R.,Cresswell,P.2009.The antiviral protein,viperin,localizes to lipid droplets via its N-terminal amphipathic-helix.Proceedings of the National Academy of Sciences 106:20452-57.
[32]Wang,H.,Ooi,E.V.,Ang,P.2008.Antiviral activities of extracts from Hong Kong seaweeds.J Zhejiang Univ Sci B 9(12):969-976.
[33]Wolf,M.C.,Freiberg,A.N.,Zhang,T.,Akyol-Ataman,Z.,Grock,A.,Hong,P.W.2010.A broad-spectrum antiviral targeting entry of enveloped viruses.Proceedings of the National Academy of Sciences 107:3157-62.
[34]Yan,R.,Zhao,Z.,He,Y.,Wu,L.,Cai,D.,Hong,W.2011.A new naturalα-helical peptide from the venom of the scorpion Heterometruspetersii kills HCV.Peptides 32:11-19.
[35]Castel,G.,Chtéoui,M.,Heyd,B.,Tordo,N.2011.Phage Display of Combinatorial Peptide Libraries:Application to Antiviral Research.Molecules 16:3499-518.
[36]Cheng,G.,Montero,A.,Gastaminza,P.,Whitten-Bauer,C.,Wieland,S.F.,Isogawa,M.2008.A virocidal amphipathic-helical peptide that inhibits hepatitis C virus infection in vitro.Proceedings of the National Academy of Sciences 105:3088-93.
[37]Li,Q.,Zhao,Z.,Zhou,D.,Chen,Y.,Hong,W.,Cao,L.2011.Virucidal activity of a scorpion venom peptide variant mucroporin-M1 against measles,SARS-Co V and influenza H5N1 viruses.Peptides 32:1518-25.
[38]You,J.H.,Howell,G.,Pattnaik,A.K.,Osorio,F.A.,Hiscox,J.A.2008.A model for the dynamic nuclear/nucleolar/cytoplasmic trafficking of the porcine reproductive and respiratory syndrome virus(PRRSV)nucleocapsid protein based on live cell imaging.Virology 378:34-47.
[2]Benfield DA,Nelson E,Collins JE,Harris L,Goyal SM,Robison D,Christianson WT,Morrison RB,Gorcyca D and Chladek D.1992.Characterization of swine infertility and respiratory syndrome(SIRS)virus(isolate ATCC VR-2332).Journal of Veterinary Diagnostic Investigation 4:127-133.
[3]Meng,X.J.,Paul,P.S.,Halbur,P.G.,Lum,M.A.1995.Phylogenetic analyses of the putative M(ORF 6)and N(ORF 7)genes of porcine reproductive and respiratory syndrome virus(PRRSV):implication for the existence of two genotypes of PRRSV in the USA and Europe.Archives of Virology 140:745-55.
[4]Mounir,S.,Mardassi,H.,Dea,S.1995.Identification and characterization of the porcine reproductive and respiratory virus ORFs 7,5 and 4 products.Advances in Experimental Medicine and Biology 380:317-20.
[5]Nelson,E.A.,Christopher-Hennings,J.,Benfield,D.A.1995.Structural proteins of porcine reproductive and respiratory syndrome virus(PRRSV).Advances in Experimental Medicine and Biology 380:321-23.
[6]Fang,Y.,Kim,D-Y.,Ropp,S.,Steen,P.,Christopher-Hennings,J.,Nelson,E.A.2004.Heterogeneity in Nsp2of European-like porcine reproductive and respiratory syndrome viruses isolated in the United States.Virus Research 100:229-35.
[7]Cafruny,W.A.,Duman,R.G.,Wong,G.H.W.,Said,S.,Ward-Demo,P.,Rowland,R.R.R.2006.Porcine reproductive and respiratory syndrome virus(PRRSV)infection spreads by cell-to-cell transfer in cultured MARC-145 cells,is dependent on an intact cytoskeleton,and is suppressed by drug-targeting of cell permissiveness to virus infection.Virology Journal 3:90.
[8]Bai,F.W.,Town,T.,Pradhan,D.,Cox,J.,Ashish,Ledizet,M.,Anderson,J.F.,Flavell,R.A.,Krueger,J.K.,Koski,R.A.2006.Antiviral Peptides Targeting the West Nile Virus Envelope Protein.Journal of Virology81:2047-55.
[9]Bajorath,J.,Peltason,L.,Wawer,M.,Guha,R.,Lajiness,M.S.,Van Drie,J.H.2009.Navigating structure-activity landscapes.Drug Discovery Today 14:698-705.
[10]Hall,P.R.,Hjelle,B.,Njus,H.,Ye,C.,Bondu-Hawkins,V.,Brown,D.C.2009.Phage Display Selection of Cyclic Peptides That Inhibit Andes Virus Infection.Journal of Virology 83:8965-69.
[11]Hancock,R.E.W.,Sahl,H-G.2006.Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies.Nature Biotechnology 24:1551-57.
[12]Ho,K.L.,Yusoff,K.,Seow,H.F.,Tan,W.S.,2003.Selection of high affinity ligands to hepatitis B core antigen from a phage-displayed cyclic peptide library.Journal of Medical Virology 69:27-32.
[13]Peng,B.,Chen,H.,Tan,Y.,Jin,M.,Guo,A.2006.Identification of one peptide which inhibited infectivity of avian infectious bronchitis virus in vitro.Science China Life Sciences 49:158-163.
[14]Sergeeva,A.,Kolonin,M.,Molldrem,J.,Pasqualini,R.,Arap,W.2006.Display technologies:Application for the discovery of drug and gene delivery agents.Advanced Drug Delivery Reviews 58:1622-54.
[15]N.Zou,J.Xia,F.Wang,Z.Duan,D.Miao,Q.Yan,S.Cao,X.Wen,P.Liu,Y.Huang.2015.Two novel neutralizing antigenic epitopes of the s1 subunit protein of a QX-like avian infectious bronchitis virus strain Sczy3as revealed using a phage display peptide library.Veterinary immunology and immunopathology,published online Epub Aug 19.
[16]Y.Du,L.Hou,J.Li.2000.Epitope mapping of HIV-1 using phage-display random peptide library and the purified Ig G from HIV patient.Chinese journal of experimental and clinical virology 14:338-341.
[17]Z.Zhang,L.Xu,Z.Wang.2011.Screening serum biomarkers for early primary hepatocellular carcinoma using a phage display technique.Journal of clinical laboratory analysis 25:402-408.
[18]Q.Deng,M.Zhuang,Y.Y.Kong,Y.H.Xie,Y.Wang.2005.Screening for Pre S specific binding ligands with a phage displayed peptides library.World journal of gastroenterology 11:4018-4023.
[19]Z.Han,F.Zhao,Y.Shao,X.Liu,X.Kong,Y.Song,S.Liu.2013.Fine level epitope mapping and conservation analysis of two novel linear B-cell epitopes of the avian infectious bronchitis coronavirus nucleocapsid protein.Virus research 171:54-64.
[20]Y.W.Zhong,D.P.Xu,X.D.Li,J.Z.Dai,B.Xu,L.Li.2009.Epitope screening of influenza A(H3N2)by using phage display library,Chinese journal of experimental and clinical virology 23:272-274.
[21]H.Xu,J.Wang,Z.Yu,F.Lv,J.Hou.2011.Screening of polypeptides binding to porcine reproductive and respiratory syndrome virus by phage display library.Wei sheng wu xue bao=Acta microbiologica Sinica51:127-133
[22]Oleksiewicz,M.B.,Botner,A.,Madsen,K.G.,Storgaard,T.1998.Sensitive detection and typing of porcine reproductive and respiratory syndrome virus by RT-PCR amplification of whole viral genes.Veterinary Microbiology 64:7-22.
[23]Sachs L,A.,Schnurr,D.,Yagi,S.,Lachowicz-Scroggins,M.E.,Widdicombe,J.H.2011.Quantitative real-time PCR for rhinovirus,and its use in determining the relationship between TCID50 and the number of viral particles.Journal of Virological Methods 171:212-18.
[24]Shimozono,M.2008.Engineering FRET Constructs Using CFP and YFP.Methods in Cell Biology85:381-393.
[25]Shyu,J.Y.,Liu,H.,Deng,X.,Hu,C.D.2006.Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions.Biotechniques 40:61-66.
[26]Mateu,E.,Diaz,I.2008.The challenge of PRRS immunology.The Veterinary Journal 177:345-51.
[27]Storgaard,T.,Oleksiewicz,M.B.,Botner,A.1999.Examination of the selective pressures on a live PRRSvaccine virus.Archives of Virology 144:2389-2401.
[28]Y.Fang,E.J.Snijder.2010.The PRRSV replicase:exploring the multifunctionality of an intriguing set of nonstructural proteins.Virus research 154:61-76.
[29]Hagiwara,K.,Kondoh,Y.,Ueda,A.,Yamada,K.,Goto,H.,Watanabe,T.2010.Discovery of novel antiviral agents directed against the influenza A virus nucleoprotein using photo-cross-linked chemical arrays.Biochemical and Biophysical Research Communications 394:721-27.
[30]Y.Sun,D.Li,S.Giri,S.G.Prasanth,D.Yoo.2014.Differential host cell gene expression and regulation of cell cycle progression by nonstructural protein 11 of porcine reproductive and respiratory syndrome virus.Bio Med research international 2014:430508
[31]Hinson,E.R.,Cresswell,P.2009.The antiviral protein,viperin,localizes to lipid droplets via its N-terminal amphipathic-helix.Proceedings of the National Academy of Sciences 106:20452-57.
[32]Wang,H.,Ooi,E.V.,Ang,P.2008.Antiviral activities of extracts from Hong Kong seaweeds.J Zhejiang Univ Sci B 9(12):969-976.
[33]Wolf,M.C.,Freiberg,A.N.,Zhang,T.,Akyol-Ataman,Z.,Grock,A.,Hong,P.W.2010.A broad-spectrum antiviral targeting entry of enveloped viruses.Proceedings of the National Academy of Sciences 107:3157-62.
[34]Yan,R.,Zhao,Z.,He,Y.,Wu,L.,Cai,D.,Hong,W.2011.A new naturalα-helical peptide from the venom of the scorpion Heterometruspetersii kills HCV.Peptides 32:11-19.
[35]Castel,G.,Chtéoui,M.,Heyd,B.,Tordo,N.2011.Phage Display of Combinatorial Peptide Libraries:Application to Antiviral Research.Molecules 16:3499-518.
[36]Cheng,G.,Montero,A.,Gastaminza,P.,Whitten-Bauer,C.,Wieland,S.F.,Isogawa,M.2008.A virocidal amphipathic-helical peptide that inhibits hepatitis C virus infection in vitro.Proceedings of the National Academy of Sciences 105:3088-93.
[37]Li,Q.,Zhao,Z.,Zhou,D.,Chen,Y.,Hong,W.,Cao,L.2011.Virucidal activity of a scorpion venom peptide variant mucroporin-M1 against measles,SARS-Co V and influenza H5N1 viruses.Peptides 32:1518-25.
[38]You,J.H.,Howell,G.,Pattnaik,A.K.,Osorio,F.A.,Hiscox,J.A.2008.A model for the dynamic nuclear/nucleolar/cytoplasmic trafficking of the porcine reproductive and respiratory syndrome virus(PRRSV)nucleocapsid protein based on live cell imaging.Virology 378:34-47.