Abstract
Protein-based nanoreactors are generated by encapsulating an enzyme inside the capsid of the cowpea chlorotic mottle virus (CCMV). Here, three different noncovalent methods are described to efficiently incorporate enzymes inside the capsid of these viral protein cages. The methods are based on pH, leucine zippers, and electrostatic interactions respectively, as a driving force for encapsulation. The methods are exclusively described for the enzymes horseradish peroxidase, glucose oxidase, and Pseudozyma antarctica lipase B, but they are also applicable for other enzymes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Minton AP, Wilf J (1981) Effect of macromolecular crowding upon the structure and function of an enzyme: glyceraldehyde-3-phosphate dehydrogenase. Biochemistry 20:4821–4826
Minton AP (2006) How can biochemical reactions within cells differ from those in test tubes? J Cell Sci 119:2863–2869
Peters RJ, Marguet M, Marais S et al (2014) Cascade reactions in multicompartmentalized polymersomes. Angew Chem Int Ed 53:146–150
Marguet M, Bonduelle C, Lecommandoux S (2013) Multicompartmentalized polymeric systems: towards biomimetic cellular structure and function. Chem Soc Rev 42:512–529
Patterson DP, Schwarz B, Waters RS et al (2014) Encapsulation of an enzyme cascade within the bacteriophage P22 virus-like particle. ACS Chem Biol 9:359–365
Patterson DP, Prevelige PE, Douglas T (2012) Nanoreactors by programmed enzyme encapsulation inside the capsid of the bacteriophage P22. ACS Nano 6:5000–5009
Minten IJ, Claessen VI, Blank K et al (2011) Catalytic capsids: the art of confinement. Chem Sci 2:358–362
Douglas T, Young M (1998) Host-guest encapsulation of materials by assembled virus protein cages. Nature 393:152–155
Bancroft JB, Hiebert E (1967) Formation of an infectious nucleoprotein from protein and nucleic acid isolated from a small spherical virus. Virology 32:354–356
Bancroft JB, Hills GJ, Markham R (1967) A study of the self-assembly process in a small spherical virus. Formation of organized structures from protein subunits in vitro. Virology 31:354–379
Speir JA, Munshi S, Wang G et al (1995) Structures of the native and swollen forms of cowpea chlorotic mottle virus determined by X-ray crystallography and cryo-electron microscopy. Structure 3:63–78
Comellas-Aragones M, Engelkamp H, Claessen VI et al (2007) A virus-based single-enzyme nanoreactor. Nat Nanotechnol 2:635–639
Kwak M, Minten IJ, Anaya DM et al (2010) Virus-like particles templated by DNA micelles: a general method for loading virus nanocarriers. J Am Chem Soc 132:7834–7835
Brasch M, de la Escosura A, Ma Y et al (2011) Encapsulation of phthalocyanine supramolecular stacks into virus-like particles. J Am Chem Soc 133:6878–6881
Minten IJ, Ma Y, Hempenius MA et al (2009) CCMV capsid formation induced by a functional negatively charged polymer. Org Biomol Chem 7:4685–4688
Comellas-Aragones M, de la Escosura A, Dirks AT et al (2009) Controlled integration of polymers into viral capsids. Biomacromolecules 10:3141–3147
Minten IJ, Hendriks LJ, Nolte RJ et al (2009) Controlled encapsulation of multiple proteins in virus capsids. J Am Chem Soc 131:17771–17773
Brasch M, Putri RM, de Ruiter MV et al (2017) Assembling enzymatic cascade pathways inside virus-based nanocages using dual-tasking nucleic acid tags. J Am Chem Soc 139:1512–1519
Rurup WF, Verbij F, Koay MST et al (2014) Predicting the loading of virus-like particles with fluorescent proteins. Biomacromolecules 15:558–563
Acknowledgment
We acknowledge financial support from the ERC Consolidator Grant (Protcage) and the Indonesia Endowment Fund for Education (LPDP).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
de Ruiter, M.V., Putri, R.M., Cornelissen, J.J.L.M. (2018). CCMV-Based Enzymatic Nanoreactors. 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_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7808-3_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7806-9
Online ISBN: 978-1-4939-7808-3
eBook Packages: Springer Protocols