Abstract
Plant viruses are emerging as versatile tools for nanotechnology applications since it is possible to modify their multivalent protein surfaces and thereby introduce and display new functionalities. In this chapter, we describe a tobacco mosaic virus (TMV) variant that exposes two selectively addressable amino acid moieties on each of its 2130 coat protein (CP) subunits. A lysine as well as a cysteine introduced at accessible sites of every CP can be modified with amino- and/or thiol-reactive chemistry such as N-hydroxysuccinimide esters (NHS ester) and maleimide containing reagents alone or simultaneously. This enables the pairwise immobilization of distinct molecules in close vicinity to each other on the TMV surface by simple standard conjugation protocols. We describe the generation of the mutations, the virus propagation and isolation as well as the dual functionalization of the TMV variant with two fluorescent dyes. The labeling is evaluated by SDS-PAGE and spectrophotometry and the degree of labeling (DOL) calculated.
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Bittner AM, Alonso JM, Górzny ML, Wege C (2013) Nanoscale science and technology with plant viruses and bacteriophages. In: Mateu MG (ed) Structure and physics of viruses: an integrated textbook, Subcellular biochemistry, vol 68. Springer Science+Business Media, Dordrecht, pp 667–702. https://doi.org/10.1007/978-94-007-6552-8_22. (ISSN: 0306-0225)
Wen AM, Steinmetz NF (2016) Design of virus-based nanomaterials for medicine, biotechnology, and energy. Chem Soc Rev 45(15):4074-4126. https://doi.org/10.1039/C5CS00287G
Mao C, Liu A, Cao B (2009) Virus-based chemical and biological sensing. Angew Chem Int Ed 48(37):6790–6810. https://doi.org/10.1002/anie.200900231
Lomonossoff GP, Evans DJ (2014) Applications of plant viruses in bionanotechnology. Plant Viral Vectors 375:61–87. https://doi.org/10.1007/82_2011_184
Lin B, Ratna B (2014) Virus hybrids as nanomaterials: methods and protocols. In: Methods in Molecular Biology, vol 1108. Humana Press, Springer, New York, NY
Singh P, Gonzalez MJ, Manchester M (2006) Viruses and their uses in nanotechnology. Drug Development Research 67(1):23–41. https://doi.org/10.1002/ddr.20064
Young M, Willits D, Uchida M, Douglas T (2008) Plant viruses as biotemplates for materials and their use in nanotechnology. Annual Review of Phytopathology 46:361–384. https://doi.org/10.1146/annurev.phyto.032508.131939
Luckanagul J, Lee LA, Nguyen QL, Sitasuwan P, Yang XM, Shazly T, Wang Q (2012) Porous alginate hydrogel functionalized with virus as three-dimensional scaffolds for bone differentiation. Biomacromolecules 13(12):3949–3958. https://doi.org/10.1021/bm301180c
Bryksin AV, Brown AC, Baksh MM, Finn MG, Barker TH (2014) Learning from nature – novel synthetic biology approaches for biomaterial design. Acta Biomaterialia 10(4):1761–1769. https://doi.org/10.1016/j.actbio.2014.01.019
Culver JN, Brown AD, Zang F, Gnerlich M, Gerasopoulos K, Ghodssi R (2015) Plant virus directed fabrication of nanoscale materials and devices. Virology 479–480:200–212. https://doi.org/10.1016/j.virol.2015.03.008
Fan XZ, Pomerantseva E, Gnerlich M, Brown A, Gerasopoulos K, McCarthy M, Culver J, Ghodssi R (2013) Tobacco mosaic virus: a biological building block for micro/nano/biosystems. J Vac Sci Technol, A 31(5):050815. https://doi.org/10.1116/1.4816584
Koch C, Eber FJ, Azucena C, Förste A, Walheim S, Schimmel T, Bittner AM, Jeske H, Gliemann H, Eiben S, Geiger FC, Wege C (2016) Novel roles for well-known players: from tobacco mosaic virus pests to enzymatically active assemblies. Beilstein J Nanotechnol 7:605–612. https://doi.org/10.3762/bjnano.7.54
Culver JN (2002) Tobacco mosaic virus assembly and disassembly: determinants in pathogenicity and resistance. Annu Rev Phytopathol 40:287–308. https://doi.org/10.1146/annurev.phyto.40.120301.102400
Caspar DLD (1963) Assembly and stability of the tobacco mosaic virus particle. Advances in Protein Chemistry 18:37–121. https://doi.org/10.1016/S0065-3233(08)60268-5
Fromm SA, Bharat TAM, Jakobi AJ, Hagen WJH, Sachse C (2015) Seeing tobacco mosaic virus through direct electron detectors. Journal of Structural Biology 189(2):87–97. https://doi.org/10.1016/j.jsb.2014.12.002
Namba K, Pattanayek R, Stubbs G (1989) Visualization of protein-nucleic acid interactions in a virus - refined structure of intact tobacco mosaic-virus at 2.9-a resolution by X-Ray fiber diffraction. Journal of Molecular Biology 208(2):307–325. https://doi.org/10.1016/0022-2836(89)90391-4
Pomerantseva E, Gerasopoulos K, Chen XY, Rubloff G, Ghodssi R (2012) Electrochemical performance of the nanostructured biotemplated V2O5 cathode for lithium-ion batteries. Journal of Power Sources 206:282–287. https://doi.org/10.1016/j.jpowsour.2012.01.127
Wu ZY, Mueller A, Degenhard S, Ruff SE, Geiger F, Bittner AM, Wege C, Krill CE (2010) Enhancing the magnetoviscosity of ferrofluids by the addition of biological nanotubes. ACS Nano 4(8):4531–4538. https://doi.org/10.1021/nn100645e
Koch C, Wabbel K, Eber FJ, Krolla-Sidenstein P, Azucena C, Gliemann H, Eiben S, Geiger F, Wege C (2015) Modified TMV particles as beneficial scaffolds to present sensor enzymes. Front Plant Sci 6:1137. https://doi.org/10.3389/fpls.2015.01137
Fan XZ, Naves L, Siwak NP, Brown A, Culver J, Ghodssi R (2015) Integration of genetically modified virus-like-particles with an optical resonator for selective bio-detection. Nanotechnology 26(20):205501. https://doi.org/10.1088/0957-4484/26/20/205501
Baecker M, Koch C, Eiben S, Geiger F, Eber FJ, Gliemann H, Poghossian A, Wege C, Schoening MJ (2017) Tobacco mosaic virus as enzyme nanocarrier for electrochemical biosensors. Sens Actuators, B 238:716–722. https://doi.org/10.1016/j.snb.2016.07.096
Bruckman MA, Randolph LN, Gulati NM, Stewart PL, Steinmetz NF (2015) Silica-coated Gd(DOTA)-loaded protein nanoparticles enable magnetic resonance imaging of macrophages. J Mater Chem B 3(38):7503–7510. https://doi.org/10.1039/C5TB01014D
Shukla S, Eber FJ, Nagarajan AS, DiFranco NA, Schmidt N, Wen AM, Eiben S, Twyman RM, Wege C, Steinmetz NF (2015) The Impact of aspect ratio on the biodistribution and tumor homing of rigid soft-matter nanorods. Adv Health Mater 4(6):874–882. https://doi.org/10.1002/adhm.201400641
Smith ML, Lindbo JA, Dillard-Telm S, Brosio PM, Lasnik AB, McCormick AA, Nguyen LV, Palmer KE (2006) Modified tobacco mosaic virus particles as scaffolds for display of protein antigens for vaccine applications. Virology 348(2):475–488. https://doi.org/10.1016/j.virol.2005.12.039
Banik S, Mansour AA, Suresh RV, Wykoff-Clary S, Malik M, McCormick AA, Bakshi CS (2015) Development of a multivalent subunit vaccine against tularemia using tobacco mosaic virus (TMV) based delivery system. Plos One 10(6):e0130858. https://doi.org/10.1371/journal.pone.0130858
Kaur G, Wang C, Sun JA, Wang QA (2010) The synergistic effects of multivalent ligand display and nanotopography on osteogenic differentiation of rat bone marrow stem cells. Biomaterials 31(22):5813–5824. https://doi.org/10.1016/j.biomaterials.2010.04.017
Zan XJ, Feng S, Balizan E, Lin Y, Wang Q (2013) Facile method for large scale alignment of one dimensional nanoparticles and control over myoblast orientation and differentiation. ACS Nano 7(10):8385–8396. https://doi.org/10.1021/nn403908k
Sitasuwan P, Lee LA, Bo P, Davis EN, Lin Y, Wang Q (2012) A plant virus substrate induces early upregulation of BMP2 for rapid bone formation. Integr Biol (Camb) 4(6):651–660. https://doi.org/10.1039/c2ib20041d
Eber FJ, Eiben S, Jeske H, Wege C (2013) Bottom-up-assembled nanostar colloids of gold cores and tubes derived from tobacco mosaic virus. Angewandte Chemie-International Edition 52(28):7203–7207. https://doi.org/10.1002/anie.201300834
Eber FJ, Eiben S, Jeske H, Wege C (2015) RNA-controlled assembly of tobacco mosaic virus-derived complex structures: from nanoboomerangs to tetrapods. Nanoscale 7(1):344–355. https://doi.org/10.1039/c4nr05434b
Taliansky ME, Kaplan IB, Yarvekulg LV, Atabekova TI, Agranovsky AA, Atabekov JG (1982) A study of TMV ts mutant Ni2519. II. Temperature-sensitive behavior of Ni2519 RNA upon reassembly. Virology 118(2):309–316. https://doi.org/10.1016/0042-6822(82)90350-6
Gallie DR, Plaskitt KA, Wilson TMA (1987) The effect of multiple dispersed copies of the origin-of-assembly sequence from TMV RNA on the morphology of pseudovirus particles assembled in vitro. Virology 158(2):473–476. https://doi.org/10.1016/0042-6822(82)90350-6
Eiben S, Stitz N, Eber F, Wagner J, Atanasova P, Bill J, Wege C, Jeske H (2014) Tailoring the surface properties of tobacco mosaic virions by the integration of bacterially expressed mutant coat protein. Virus Research 180:92–96. https://doi.org/10.1016/j.virusres.2013.11.019
Geiger FC, Eber FJ, Eiben S, Mueller A, Jeske H, Spatz JP, Wege C (2013) TMV nanorods with programmed longitudinal domains of differently addressable coat proteins. Nanoscale 5(9):3808–3816. https://doi.org/10.1039/c3nr33724c
Schneider A, Eber FJ, Wenz N, Altintoprak K, Jeske H, Eiben S, Wege C (2016) Dynamic DNA-controlled “stop-and-go” assembly of well-defined protein domains on RNA-scaffolded TMV-like nanotubes. Nanoscale 8:19853–19866. https://doi.org/10.1039/C6NR03897B
Royston E, Ghosh A, Kofinas P, Harris MT, Culver JN (2008) Self-assembly of virus-structured high surface area nanomaterials and their application as battery electrodes. Langmuir 24(3):906–912. https://doi.org/10.1021/la7016424
Yi H, Rubloff GW, Culver JN (2007) TMV microarrays: hybridization-based assembly of DNA-programmed viral nanotemplates. Langmuir 23(5):2663–2667. https://doi.org/10.1021/la062493c
Mueller A, Eber FJ, Azucena C, Petershans A, Bittner AM, Gliemann H, Jeske H, Wege C (2011) Inducible site-selective bottom-up assembly of virus-derived nanotube arrays on RNA-equipped wafers. ACS Nano 5(6):4512–4520. https://doi.org/10.1021/nn103557s
Butler PJ (1999) Self-assembly of tobacco mosaic virus: the role of an intermediate aggregate in generating both specificity and speed. Philos Trans R Soc Lond B Biol Sci 354(1383):537–550. https://doi.org/10.1098/rstb.1999.0405
Bruckman MA, Steinmetz NF (2014) Chemical modification of the inner and outer surfaces of tobacco mosaic virus (TMV). Virus Hybrids as Nanomaterials: Methods and Protocols 1108:173–185. https://doi.org/10.1007/978-1-62703-751-8_13
Bruckman MA, Kaur G, Lee LA, Xie F, Sepulveda J, Breitenkamp R, Zhang X, Joralemon M, Russell TP, Emrick T, Wang Q (2008) Surface modification of tobacco mosaic virus with “click” chemistry. Chembiochem 9(4):519–523. https://doi.org/10.1002/cbic.200700559
Wu LY, Zang JF, Lee LA, Niu ZW, Horvatha GC, Braxtona V, Wibowo AC, Bruckman MA, Ghoshroy S, zur Loye HC, Li XD, Wang Q (2011) Electrospinning fabrication, structural and mechanical characterization of rod-like virus-based composite nanofibers. Journal of Materials Chemistry 21(24):8550–8557. https://doi.org/10.1039/c1jm00078k
Witus LS, Francis MB (2011) Using synthetically modified proteins to make new materials. Acc Chem Res 44(9):774–783. https://doi.org/10.1098/rstb.1999.0405
McKay Craig S, Finn MG (2014) Click chemistry in complex mixtures: bioorthogonal bioconjugation. Chemistry & Biology 21(9):1075–1101. https://doi.org/10.1016/j.chembiol.2014.09.002
Wu FC, Zhang H, Zhou Q, Wu M, Ballard Z, Tian Y, Wang JY, Niu ZW, Huang Y (2014) Expanding the genetic code for site-specific labelling of tobacco mosaic virus coat protein and building biotin-functionalized virus-like particles. Chem Commun 50(30):4007–4009. https://doi.org/10.1039/c3cc49137d
Zhou K, Li F, Dai GL, Meng C, Wang QB (2013) Disulfide bond: dramatically enhanced assembly capability and structural stability of tobacco mosaic virus nanorods. Biomacromolecules 14(8):2593–2600. https://doi.org/10.1021/bm400445m
Miller RA, Presley AD, Francis MB (2007) Self-assembling light-harvesting systems from synthetically modified tobacco mosaic virus coat proteins. Journal of the American Chemical Society 129(11):3104–3109. https://doi.org/10.1021/ja063887t
Demir M, Stowell MHB (2002) A chemoselective biomolecular template for assembling diverse nanotubular materials. Nanotechnology 13(4):541–544. https://doi.org/10.1088/0957-4484/13/4/318. Pii S0957-4484(02)31463-6
Yi HM, Nisar S, Lee SY, Powers MA, Bentley WE, Payne GF, Ghodssi R, Rubloff GW, Harris MT, Culver JN (2005) Patterned assembly of genetically modified viral nanotemplates via nucleic acid hybridization. Nano Letters 5(10):1931–1936. https://doi.org/10.1021/nl051254r
Lee SY, Choi J, Royston E, Janes DB, Culver JN, Harris MT (2006) Deposition of platinum clusters on surface-modified tobacco mosaic virus. J Nanosci Nanotechnol 6(4):974–981. https://doi.org/10.1098/rstb.1999.0405
Lim JS, Kim SM, Lee SY, Stach EA, Culver JN, Harris MT (2010) Biotemplated aqueous-phase palladium crystallization in the absence of external reducing agents. Nano Lett 10(10):3863–3867. https://doi.org/10.1021/nl101375f
Yang C, Choi CH, Lee CS, Yi H (2013) A facile synthesis-fabrication strategy for integration of catalytically active viral-palladium nanostructures into polymeric hydrogel microparticles via replica molding. ACS Nano 7(6):5032–5044. https://doi.org/10.1021/nn4005582
Harder A, Dieding M, Walhorn V, Degenhard S, Brodehl A, Wege C, Milting H, Anselmetti D (2013) Apertureless scanning near-field optical microscopy of sparsely labeled tobacco mosaic viruses and the intermediate filament desmin. Beilstein J Nanotechnol 4:510–516. https://doi.org/10.3762/bjnano.4.60
Altintoprak K, Seidenstucker A, Welle A, Eiben S, Atanasova P, Stitz N, Plettl A, Bill J, Gliemann H, Jeske H, Rothenstein D, Geiger F, Wege C (2015) Peptide-equipped tobacco mosaic virus templates for selective and controllable biomineral deposition. Beilstein Journal of Nanotechnology 6:1399–1412. https://doi.org/10.3762/bjnano.6.145
Kadri A, Maiss E, Amsharov N, Bittner AM, Balci S, Kern K, Jeske H, Wege C (2011) Engineered tobacco mosaic virus mutants with distinct physical characteristics in planta and enhanced metallization properties. Virus Res 157(1):35–46. https://doi.org/10.1016/j.virusres.2011.01.014
Gooding GV Jr, Hebert TT (1967) A simple technique for purification of tobacco mosaic virus in large quantities. Phytopathology 57(11):1285
Green MR, Sambrook J (2012) In: Inglis J (ed) Molecular cloning a laboratory manual, vol 1, 4th edn. Cold Spring Harbor, New York, NY
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Wege, C., Geiger, F. (2018). Dual Functionalization of Rod-Shaped Viruses on Single Coat Protein Subunits. In: Wege, C., Lomonossoff, G. (eds) Virus-Derived Nanoparticles for Advanced Technologies. Methods in Molecular Biology, vol 1776. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7808-3_27
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