埃博拉病毒糖蛋白与宿主细胞相互作用蛋白的筛选及生物信息学分析
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摘要
目的筛选埃博拉病毒糖蛋白在人源宿主细胞内的相互作用组分,为埃博拉病毒受体研究提供基础数据。方法制备埃博拉病毒糖蛋白GP1亚基人IgG-Fc标签融合蛋白,进行Pull-Down试验和质谱分析,经生物信息学分析确定关键相互作用因子。结果共筛选出31种潜在的与宿主细胞相互助用的埃博拉病毒糖蛋白,该类蛋白或为细胞结构成分,或为转录因子并参与信号转导等生物学过程;有10种蛋白参与20种信号通路,主要包括免疫反应、细胞黏附、致病性、病原感染等4大类信号通路。结论成功筛选与人源细胞相互作用的埃博拉病毒糖蛋白,其中部分蛋白在细胞生命活动中起重要作用。
Objective To screen for proteins interacting with Ebola virus(EBOV)glycoprotein(GP)in human host cells and to provide basic data for research on EBOV receptors. Methods The EBOV glycoprotein subunit 1(GP1)fusion protein with human IgG-Fc,named GP1-Fc,was used to perform a pull-down experiment.Components interacting with GP1 in A549 cells were identified using mass spectrometry and bioinformatic analysis. Results Thirty-one potentially interacting proteins were identified.These proteins might play important roles in many biological functions such as cellular structure,transcription,and signaling.Ten proteins were found to be involved in 20 signaling pathways which were mainly 4 types of pathways:immunoreaction,cell adhesion,pathogenicity and adventitious infection.Using pulldown biotechnology to examine interacting components and using GO and Pathway analysis to bioinformatically analyze identified proteins might prove of use in future studies of EBOV GP. Conclusion This study successfully identified proteins that potentially interact with EBOV GP in human host cells.These proteins might play important roles in many cellular biological activities.
Objective To screen for proteins interacting with Ebola virus(EBOV)glycoprotein(GP)in human host cells and to provide basic data for research on EBOV receptors. Methods The EBOV glycoprotein subunit 1(GP1)fusion protein with human IgG-Fc,named GP1-Fc,was used to perform a pull-down experiment.Components interacting with GP1 in A549 cells were identified using mass spectrometry and bioinformatic analysis. Results Thirty-one potentially interacting proteins were identified.These proteins might play important roles in many biological functions such as cellular structure,transcription,and signaling.Ten proteins were found to be involved in 20 signaling pathways which were mainly 4 types of pathways:immunoreaction,cell adhesion,pathogenicity and adventitious infection.Using pulldown biotechnology to examine interacting components and using GO and Pathway analysis to bioinformatically analyze identified proteins might prove of use in future studies of EBOV GP. Conclusion This study successfully identified proteins that potentially interact with EBOV GP in human host cells.These proteins might play important roles in many cellular biological activities.
引文
[1]Becker S,Spiess M,Klenk HD.The asialoglycoprotein receptor is a potential liver-specific receptor for Marburg virus[J].J Gen Virol,1995,76(Pt 2):393-9.
[2]Chan SY,Empig CJ,Welte FJ,et al.Folate receptor-alpha is a cofactor for cellular entry by Marburg and Ebola viruses[J].Cell,2001,106(1):117-26.
[3]Alvarez CP,Lasala F,Carrillo J,et al.C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans[J].J Virol,2002,76(13):6841-4.
[4]Takada A,Fujioka K,Tsuiji M,et al.Human macrophage Ctype lectin specific for galactose and N-acetylgalactosamine promotes filovirus entry[J].J Virol,2004,78(6):2943-7.
[5]Chandran K,Sullivan NJ,Felbor U,et al.Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection[J].Science,2005,308(5728):1643-5.
[6]Shimojima M,Takada A,Ebihara H,et al.Tyro3family-mediated cell entry of Ebola and Marburg viruses[J].J Virol,2006,80(20):10109-116.
[7]Kondratowicz AS,Lennemann NJ,Sinn PL,et al.T-cell immunoglobulin and mucin domain 1(TIM-1)is a receptor for Zaire Ebolavirus and Lake Victoria Marburgvirus[J].Proc Natl Acad Sci USA,2011,108(20):8426-31.
[8]Carette JE,Raaben M,Wong AC,et al.Ebola virus entry requires the cholesterol transporter Niemann-Pick C1[J].Nature,2011,477(7364):340-3.
[9]Cote M,Misasi J,Ren T,et al.Small molecule inhibitors reveal Niemann-Pick C1is essential for Ebola virus infection[J].Nature,2011,477(7364):344-8.
[10]Miller EH,Obernosterer G,Raaben M,et al.Ebola virus entry requires the host-programmed recognition of an intracellular receptor[J].EMBO J,2012,31(8):1947-60.
[11]Salvador B,Sexton NR,Carrion R,et al.Filoviruses utilize glycosaminoglycans for their attachment to target cells[J].J Virol,2013,87(6):3295-304.
[12]O'hearn A,Wang M,Cheng H,et al.Role of EXT1and Glycosaminoglycans in the Early Stage of Filovirus Entry[J].J Virol,2015,89(10):5441-9.
[13]Kuroda M,Fujikura D,Nanbo A,et al.Interaction between TIM-1and NPC1is important for cellular entry of Ebola virus[J].J Virol,2015,89(12):6481-93.
[14]王中一,李佳明,付莹莹,等.埃博拉病毒糖蛋白GP1亚基融合蛋白的真核表达与纯化[J].吉林农业大学学报,2018,(04):1-6.
[15]Wang ZY,Guo ZD,Li J M,et al.Genome-wide search for competing endogenous RNAs responsible for the effects induced by Ebola virus replication and transcription using a trVLP system[J].Front Cell Infect Microbiol,2017(7):479.
[16]蔡建秋,徐琳,涂长春,等.埃博拉病毒核蛋白抗体间接ELISA检测方法的建立[J].中国病原生物学杂志,2017,12(1):1-4,9.
[17]Twenhafel NA,Mattix ME,Johnson JC,et al.Pathology of experimental aerosol Zaire ebolavirus infection in rhesus macaques[J].Vet Pathol,2013,50(3):514-9.
[18]Gregory SG,Barlow KF,Mclay KE,et al.The DNA sequence and biological annotation of human chromosome 1[J].Nature,2006,441(7091):315-21.
[19]Burgisser DM,Siegenthaler G,Kuster T,et al.Amino acid sequence analysis of human S100A7(psoriasin)by tandem mass spectrometry[J].Biochem Biophys Res Commun,1995,217(1):257-63.
[20]Fagerberg L,Hallstrom BM,Oksvold P,et al.Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics[J].Mol Cell Proteomics,2014,13(2):397-406.
[21]Wu Z,Hansmann B,Meyer-Hoffert U,et al.Molecular identification and expression analysis of filaggrin-2,a member of the S100fused-type protein family[J].PLoS One,2009,4(4):e5227.
[2]Chan SY,Empig CJ,Welte FJ,et al.Folate receptor-alpha is a cofactor for cellular entry by Marburg and Ebola viruses[J].Cell,2001,106(1):117-26.
[3]Alvarez CP,Lasala F,Carrillo J,et al.C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans[J].J Virol,2002,76(13):6841-4.
[4]Takada A,Fujioka K,Tsuiji M,et al.Human macrophage Ctype lectin specific for galactose and N-acetylgalactosamine promotes filovirus entry[J].J Virol,2004,78(6):2943-7.
[5]Chandran K,Sullivan NJ,Felbor U,et al.Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection[J].Science,2005,308(5728):1643-5.
[6]Shimojima M,Takada A,Ebihara H,et al.Tyro3family-mediated cell entry of Ebola and Marburg viruses[J].J Virol,2006,80(20):10109-116.
[7]Kondratowicz AS,Lennemann NJ,Sinn PL,et al.T-cell immunoglobulin and mucin domain 1(TIM-1)is a receptor for Zaire Ebolavirus and Lake Victoria Marburgvirus[J].Proc Natl Acad Sci USA,2011,108(20):8426-31.
[8]Carette JE,Raaben M,Wong AC,et al.Ebola virus entry requires the cholesterol transporter Niemann-Pick C1[J].Nature,2011,477(7364):340-3.
[9]Cote M,Misasi J,Ren T,et al.Small molecule inhibitors reveal Niemann-Pick C1is essential for Ebola virus infection[J].Nature,2011,477(7364):344-8.
[10]Miller EH,Obernosterer G,Raaben M,et al.Ebola virus entry requires the host-programmed recognition of an intracellular receptor[J].EMBO J,2012,31(8):1947-60.
[11]Salvador B,Sexton NR,Carrion R,et al.Filoviruses utilize glycosaminoglycans for their attachment to target cells[J].J Virol,2013,87(6):3295-304.
[12]O'hearn A,Wang M,Cheng H,et al.Role of EXT1and Glycosaminoglycans in the Early Stage of Filovirus Entry[J].J Virol,2015,89(10):5441-9.
[13]Kuroda M,Fujikura D,Nanbo A,et al.Interaction between TIM-1and NPC1is important for cellular entry of Ebola virus[J].J Virol,2015,89(12):6481-93.
[14]王中一,李佳明,付莹莹,等.埃博拉病毒糖蛋白GP1亚基融合蛋白的真核表达与纯化[J].吉林农业大学学报,2018,(04):1-6.
[15]Wang ZY,Guo ZD,Li J M,et al.Genome-wide search for competing endogenous RNAs responsible for the effects induced by Ebola virus replication and transcription using a trVLP system[J].Front Cell Infect Microbiol,2017(7):479.
[16]蔡建秋,徐琳,涂长春,等.埃博拉病毒核蛋白抗体间接ELISA检测方法的建立[J].中国病原生物学杂志,2017,12(1):1-4,9.
[17]Twenhafel NA,Mattix ME,Johnson JC,et al.Pathology of experimental aerosol Zaire ebolavirus infection in rhesus macaques[J].Vet Pathol,2013,50(3):514-9.
[18]Gregory SG,Barlow KF,Mclay KE,et al.The DNA sequence and biological annotation of human chromosome 1[J].Nature,2006,441(7091):315-21.
[19]Burgisser DM,Siegenthaler G,Kuster T,et al.Amino acid sequence analysis of human S100A7(psoriasin)by tandem mass spectrometry[J].Biochem Biophys Res Commun,1995,217(1):257-63.
[20]Fagerberg L,Hallstrom BM,Oksvold P,et al.Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics[J].Mol Cell Proteomics,2014,13(2):397-406.
[21]Wu Z,Hansmann B,Meyer-Hoffert U,et al.Molecular identification and expression analysis of filaggrin-2,a member of the S100fused-type protein family[J].PLoS One,2009,4(4):e5227.