Hostname: page-component-cd9895bd7-dk4vv Total loading time: 0 Render date: 2024-12-27T09:27:03.899Z Has data issue: false hasContentIssue false
  • English
  • Français

Capturing Enveloped Viruses on Affinity Grids for Downstream Cryo-Electron Microscopy Applications

Published online by Cambridge University Press:  26 November 2013

Gabriella Kiss ,
Xuemin Chen ,
Melinda A. Brindley ,
Patricia Campbell ,
Claudio L. Afonso ,
Zunlong Ke ,
Jens M. Holl ,
Ricardo C. Guerrero-Ferreira ,
Lauren A. Byrd-Leotis  and
John Steel
...Show all authors
Gabriella Kiss
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA
Xuemin Chen
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA
Melinda A. Brindley
Affiliation:
Center for Inflammation, Immunity & Infection, Georgia State University, Atlanta, GA 30303, USA
Patricia Campbell
Affiliation:
Department of Microbiology and Immunology, School of Medicine, Emory University, Atlanta, GA 30322, USA
Claudio L. Afonso
Affiliation:
USDA, ARS, Southeast Poultry Research Laboratory, Athens, GA 30605, USA
Zunlong Ke
Affiliation:
School of Biology, Georgia Institute of Technology, Atlanta, GA 30332, USA
Jens M. Holl
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA
Ricardo C. Guerrero-Ferreira
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA
Lauren A. Byrd-Leotis
Affiliation:
Department of Microbiology and Immunology, School of Medicine, Emory University, Atlanta, GA 30322, USA
John Steel
Affiliation:
Department of Microbiology and Immunology, School of Medicine, Emory University, Atlanta, GA 30322, USA
David A. Steinhauer
Affiliation:
Department of Microbiology and Immunology, School of Medicine, Emory University, Atlanta, GA 30322, USA
Richard K. Plemper
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA Center for Inflammation, Immunity & Infection, Georgia State University, Atlanta, GA 30303, USA
Deborah F. Kelly
Affiliation:
Virginia Tech Carilion Research Institute, Roanoke, VA 24016, USA
Paul W. Spearman
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA
Elizabeth R. Wright*
Affiliation:
Department of Pediatrics, School of Medicine, Division of Pediatric Infectious Diseases, Emory University, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA
*
*Corresponding author. E-mail: [email protected]
Get access

Abstract

Electron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.

Keywords

cryo-electron microscopy (cryo-EM)cryo-electron tomography (cryo-ET)affinity grid (AG)enveloped virus
Type
Techniques, Software, and Instrumentation Development
Information
Microscopy and Microanalysis , Volume 20 , Issue 1 , February 2014 , pp. 164 - 174
Copyright
Copyright © Microscopy Society of America 2014 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Briggs, J.A., Grunewald, K., Glass, B., Forster, F., Krausslich, H.G. & Fuller, S.D. (2006a). The mechanism of HIV-1 core assembly: Insights from three-dimensional reconstructions of authentic virions. Structure 14, 15–20.Google Scholar
Briggs, J.A., Johnson, M.C., Simon, M.N., Fuller, S.D. & Vogt, V.M. (2006b). Cryo-electron microscopy reveals conserved and divergent features of Gag packing in immature particles of Rous sarcoma virus and human immunodeficiency virus. J Mol Biol 355, 157–168.Google Scholar
Butan, C., Winkler, D.C., Heymann, J.B., Craven, R.C. & Steven, A.C. (2008). RSV capsid polymorphism correlates with polymerization efficiency and envelope glycoprotein content: Implications that nucleation controls morphogenesis. J Mol Biol 376, 1168–1181.Google Scholar
Calder, L.J., Wasilewski, S., Berriman, J.A. & Rosenthal, P.B. (2010). Structural organization of a filamentous influenza A virus. Proc Natl Acad Sci USA 107, 10685–10690.Google Scholar
Carlson, L.A., Briggs, J.A., Glass, B., Riches, J.D., Simon, M.N., Johnson, M.C., Muller, B., Grunewald, K. & Krausslich, H.G. (2008). Three-dimensional analysis of budding sites and released virus suggests a revised model for HIV-1 morphogenesis. Cell Host Microbe 4, 592–599.Google Scholar
de Marco, A., Davey, N.E., Ulbrich, P., Phillips, J.M., Lux, V., Riches, J.D., Fuzik, T., Ruml, T., Krausslich, H.G., Vogt, V.M. & Briggs, J.A. (2010). Conserved and variable features of Gag structure and arrangement in immature retrovirus particles. J Virol 84, 11729–11736.Google Scholar
Fodor, E., Devenish, L., Engelhardt, O.G., Palese, P., Brownlee, G.G. & Garcia-Sastre, A. (1999). Rescue of influenza A virus from recombinant DNA. J Virol 73, 9679–9682.Google Scholar
Fontana, J., Cardone, G., Heymann, J.B., Winkler, D.C. & Steven, A.C. (2012). Structural changes in influenza virus at low pH characterized by cryo-electron tomography. J Virol 86, 2919–2929.Google Scholar
Gias, E., Nielsen, S.U., Morgan, L.A. & Toms, G.L. (2008). Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient. J Virol Methods 147, 328–332.Google Scholar
Gilmore, B.L., Showalter, S.P., Dukes, M.J., Tanner, J.R., Demmert, A.C., McDonald, S.M. & Kelly, D.F. (2013). Visualizing viral assemblies in a nanoscale biosphere. Lab Chip 13, 216–219.Google Scholar
Hall, W.W., Martin, S.J. & Gould, E. (1974). Defective interfering particles produced during the replication of measles virus. Med Microbiol Immunol 160, 155–164.CrossRefGoogle ScholarPubMed
Hammonds, J., Chen, X., Zhang, X., Lee, F. & Spearman, P. (2007). Advances in methods for the production, purification, and characterization of HIV-1 Gag-Env pseudovirion vaccines. Vaccine 25, 8036–8048.Google Scholar
Harris, A., Cardone, G., Winkler, D.C., Heymann, J.B., Brecher, M., White, J.M. & Steven, A.C. (2006). Influenza virus pleiomorphy characterized by cryoelectron tomography. Proc Natl Acad Sci USA 103, 19123–19127.Google Scholar
Heymann, J.B., Butan, C., Winkler, D.C., Craven, R.C. & Steven, A.C. (2008). Irregular and semi-regular polyhedral models for Rous sarcoma virus cores. Comput Math Methods Med 9, 197–210.CrossRefGoogle ScholarPubMed
Hicar, M.D., Chen, X., Briney, B., Hammonds, J., Wang, J.J., Kalams, S., Spearman, P.W. & Crowe, J.E. Jr. (2010). Pseudovirion particles bearing native HIV envelope trimers facilitate a novel method for generating human neutralizing monoclonal antibodies against HIV. J Acquir Immune Defic Syndr 54, 223–235.Google Scholar
Keller, P.W., Adamson, C.S., Heymann, J.B., Freed, E.O. & Steven, A.C. (2011). HIV-1 maturation inhibitor bevirimat stabilizes the immature Gag lattice. J Virol 85, 1420–1428.CrossRefGoogle ScholarPubMed
Kelly, D.F., Abeyrathne, P.D., Dukovski, D. & Walz, T. (2008a). The affinity grid: A pre-fabricated EM grid for monolayer purification. J Mol Biol 382, 423–433.Google Scholar
Kelly, D.F., Dukovski, D. & Walz, T. (2008b). Monolayer purification: A rapid method for isolating protein complexes for single-particle electron microscopy. Proc Natl Acad Sci USA 105, 4703–4708.Google Scholar
Kelly, D.F., Dukovski, D. & Walz, T. (2010a). A practical guide to the use of monolayer purification and affinity grids. Methods Enzymol 481, 83–107.Google Scholar
Kelly, D.F., Dukovski, D. & Walz, T. (2010b). Strategy for the use of affinity grids to prepare non-His-tagged macromolecular complexes for single-particle electron microscopy. J Mol Biol 400, 675–681.Google Scholar
Kremer, J.R., Mastronarde, D.N. & McIntosh, J.R. (1996). Computer visualization of three-dimensional image data using IMOD. J Struct Biol 116, 71–76.CrossRefGoogle ScholarPubMed
Lee, K.K. (2010). Architecture of a nascent viral fusion pore. EMBO J 29, 1299–1311.Google Scholar
Liljeroos, L., Huiskonen, J.T., Ora, A., Susi, P. & Butcher, S.J. (2011). Electron cryotomography of measles virus reveals how matrix protein coats the ribonucleocapsid within intact virions. Proc Natl Acad Sci USA 108, 18085–18090.Google Scholar
Liljeroos, L., Krzyzaniak, M.A., Helenius, A. & Butcher, S.J. (2013). Architecture of respiratory syncytial virus revealed by electron cryotomography. Proc Natl Acad Sci USA 110, 11133–11138.Google Scholar
Loney, C., Mottet-Osman, G., Roux, L. & Bhella, D. (2009). Paramyxovirus ultrastructure and genome packaging: Cryo-electron tomography of sendai virus. J Virol 83, 8191–8197.Google Scholar
Maeda, A., Suzuki, Y. & Matsumoto, M. (1978). Isolation and characterization of defective interfering particle of Newcastle disease virus. Microbiol Immunol 22, 775–784.Google Scholar
Mastronarde, D.N. (2005). Automated electron microscope tomography using robust prediction of specimen movements. J Struct Biol 152, 36–51.Google Scholar
Mbiguino, A. & Menezes, J. (1991). Purification of human respiratory syncytial virus: Superiority of sucrose gradient over percoll, renografin, and metrizamide gradients. J Virol Methods 31, 161–170.CrossRefGoogle ScholarPubMed
McGinnes, L.W., Pantua, H., Reitter, J. & Morrison, T.G. (2006). Newcastle disease virus: Propagation, quantification, and storage. Curr Protoc Microbiol Chapter 15, Unit 15F, 12.Google Scholar
Ono, N., Tatsuo, H., Hidaka, Y., Aoki, T., Minagawa, H. & Yanagi, Y. (2001). Measles viruses on throat swabs from measles patients use signaling lymphocytic activation molecule (CDw150) but not CD46 as a cellular receptor. J Virol 75, 4399–4401.Google Scholar
Plemper, R.K., Hammond, A.L. & Cattaneo, R. (2001). Measles virus envelope glycoproteins hetero-oligomerize in the endoplasmic reticulum. J Biol Chem 276, 44239–44246.Google Scholar
Sugita, Y., Noda, T., Sagara, H. & Kawaoka, Y. (2011). Ultracentrifugation deforms unfixed influenza A virions. J Gen Virol 92, 2485–2493.Google Scholar
Tanner, J.R., Degen, K., Gilmore, B.L. & Kelly, D.F. (2012). Capturing RNA-dependent pathways for cryo-EM analysis. Comput Struct Biotech J 1, e201204003-1–6.Google Scholar
Valdovinos, M.R. & Gomez, B. (2003). Establishment of respiratory syncytial virus persistence in cell lines: Association with defective interfering particles. Intervirology 46, 190–198.Google Scholar
Wickramasinghe, S.R., Kalbfuss, B., Zimmermann, A., Thom, V. & Reichl, U. (2005). Tangential flow microfiltration and ultrafiltration for human influenza A virus concentration and purification. Biotechnol Bioeng 92, 199–208.Google Scholar
Wright, E.R., Schooler, J.B., Ding, H.J., Kieffer, C., Fillmore, C., Sundquist, W.I. & Jensen, G.J. (2007). Electron cryotomography of immature HIV-1 virions reveals the structure of the CA and SP1 Gag shells. EMBO J 26, 2218–2226.Google Scholar
Yamaguchi, M., Danev, R., Nishiyama, K., Sugawara, K. & Nagayama, K. (2008). Zernike phase contrast electron microscopy of ice-embedded influenza A virus. J Struct Biol 162, 271–276.Google Scholar
Zhang, L., Hernan, R. & Brizzard, B. (2001). Multiple tandem epitope tagging for enhanced detection of protein expressed in mammalian cells. Mol Biotechnol 19, 313–321.Google Scholar
Supplementary material: PDF

Kiss et al. Supplementary Material

Figures and Tables

Download Kiss et al. Supplementary Material(PDF)
PDF 1.4 MB