Implementation of P22 Viral Capsids as Nanoplatforms

Kang, Sebyung; Uchida, Masaki; O’Neil, Alison; Li, Rui; Prevelige, Peter E.; Douglas, Trevor

doi:10.1021/bm100877q

Cited by 90 publications

(133 citation statements)

References 19 publications

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“…The CP of P22 does not possess reactive cysteines 55 whereas SP-KivD and SP-AdhA have two and ten cysteine residues, respectively, hence the dyes were conjugated with the SP-enzyme fusion proteins encapsulated inside of VLPs. For P22-E2-KivD, the VLP was treated with 1 mM DTT at room temperature for 4h prior to the labeling with DyLight 405.…”

Section: Methodsmentioning

confidence: 99%

Modular Self-Assembly of Protein Cage Lattices for Multistep Catalysis

Uchida

McCoy

Fukuto³

et al. 2017

ACS Nano

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190

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The assembly of individual molecules into hierarchical structures is a promising strategy for developing three-dimensional materials with properties arising from interaction between the individual building blocks. Virus capsids are elegant examples of biomolecular nanostructures, which are themselves hierarchically assembled from a limited number of protein subunits. Here we demonstrate the bio-inspired modular construction of materials with two levels of hierarchy; the formation of catalytically active individual virus-like particles (VLPs) through directed self-assembly of capsid subunits with enzyme encapsulation, and the assembly of these VLP building blocks into three-dimensional arrays. The structure of the assembled arrays was successfully altered from an amorphous aggregate to an ordered structure, with a face-centered cubic lattice, by modifying the exterior surface of the VLP without changing its overall morphology, to modulate interparticle interactions. The assembly behavior and resultant lattice structure was a consequence of interparticle interaction between exterior surfaces of individual particles, and thus independent of the enzyme cargos encapsulated within the VLPs. These superlattice materials, composed of two populations of enzyme packaged VLP modules, retained the coupled catalytic activity in a two-step reaction for isobutanol synthesis. This study demonstrates a significant step toward the bottom-up fabrication of functional superlattice materials using a self-assembly process across multiple length scales, and exhibits properties and function that arise from the interaction between individual building blocks.

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Section: Methodsmentioning

confidence: 99%

Modular Self-Assembly of Protein Cage Lattices for Multistep Catalysis

Uchida

McCoy

Fukuto³

et al. 2017

ACS Nano

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190

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“…Genetic manipulation, recombinant expression, self-assembly, and surface morphology of P22 are well-established, [38] making it an excellent candidate for use in nanotechnology. When expressed in E. coli, in the presence of scaffolding proteins of P22, the P22 coat proteins assemble into an icosahedral T = 7 procapsid structure comprising 420 identical subunits.…”

Section: Introductionmentioning

confidence: 99%

Virus Matryoshka: A Bacteriophage Particle—Guided Molecular Assembly Approach to a Monodisperse Model of the Immature Human Immunodeficiency Virus

Saxena

Malyutin

et al. 2016

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Immature human immunodeficiency virus type 1 (HIV-1) is approximately spherical, but is constructed from a hexagonal lattice of the Gag protein. As a hexagonal lattice is necessarily flat, the local symmetry cannot be maintained throughout the structure. This geometrical frustration presumably results in bending stress. In natural particles, the stress is relieved by incorporation of packing defects, but the magnitude of this stress and its significance for the particles is not known. In order to control this stress, we have now assembled the Gag protein on a quasi-spherical template derived from bacteriophage P22. This template is monodisperse in size and electron-transparent, enabling the use of cryo-electron microscopy in structural studies. These templated assemblies are far less polydisperse than any previously described virus-like particles (and, while constructed according to the same lattice as natural particles, contain almost no packing defects). This system gives us the ability to study the relationship between packing defects, curvature and elastic energy, and thermodynamic stability. As Gag is bound to the P22 template by single-stranded DNA, treatment of the particles with DNase enabled us to determine the intrinsic radius of curvature of a Gag lattice, unconstrained by DNA or a template. We found that this intrinsic radius is far larger than that of a virion or P22-templated particle. We conclude that Gag is under elastic strain in a particle; this has important implications for the kinetics of shell growth, the stability of the shell, and the type of defects it will assume as it grows.

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“…The HK97 fold has been detected in every tailed phage capsid that has been imaged at sufficient resolution, including P22 11; 12; 13 , T4 14 , λ 15 , ϕ29 16 , T5 17; 18 , P2/P4 19 , epsilon15 20 , T7 21; 22 , the tailed-phage-like archaevirus HSTV-1 23 , in the base layer of the Herpesvirus (HSV1) major capsid protein 24 and in the bacterial comparments known as encapsulins 25 . With renewed interest in phage therapy to combat multiple-drug-resistant bacterial pathogens 26; 27; 28 , and an emerging interest in using bacteriophage capsids as platforms for nanotechnology 29 (http://www.nano.gov/) directed to the treatment of disease 30; 31 and the creation of new technologies 32; 33; 34; 35 , it is important to have a deep understanding of capsid structures and assembly pathways in order that they can be manipulated and utilized to the greatest extent.…”

Section: Introductionmentioning

confidence: 99%

Transient Contacts on the Exterior of the HK97 Procapsid That Are Essential for Capsid Assembly

Tso¹,

Hendrix²,

Duda³

2014

Journal of Molecular Biology

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The G-loop is a 10-residue glycine-rich loop that protrudes from the surface of the mature bacteriophage HK97 capsid at the C-terminal end of the long backbone helix of major capsid protein subunits. The G-loop is essential for assembly, is conserved in related capsid and encapsulin proteins, and plays its role during HK97 capsid assembly by making crucial contacts between the hill-like hexamers and pentamers in precursor proheads. These contacts are not preserved in the flattened capsomers of the mature capsid. Aspartate 231 in each of the ~400 G-loops interacts with Lysine 178 of the E-loop (Extended Loop) of a subunit on an adjacent capsomer. Mutations disrupting this interaction prevented correct assembly, and in some cases induced abnormal assembly into tubes, or small, incomplete capsids. Assembly remained defective when D231 and K178 were replaced with larger charged residues or when their positions were exchanged. Second-site suppressors of lethal mutants containing substitution D231L replaced the ionic interaction with new interactions between neutral or hydrophobic residues of about the same size: D231L/K178V, D231L/K178I, and D231L/K178N. We conclude that it is not the charge, but the size and shape of the side chains of residues 178 and 231 that are important. These two residues control the geometry of contacts between the E-loop and the G-loop, which apparently must be precisely spaced and oriented for correct assembly to occur. We present a model for how the G-loop could control HK97 assembly and identify G-loop-like protrusions in other capsid proteins that may play analogous roles.

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Implementation of P22 Viral Capsids as Nanoplatforms

Cited by 90 publications

References 19 publications

Modular Self-Assembly of Protein Cage Lattices for Multistep Catalysis

Modular Self-Assembly of Protein Cage Lattices for Multistep Catalysis

Virus Matryoshka: A Bacteriophage Particle—Guided Molecular Assembly Approach to a Monodisperse Model of the Immature Human Immunodeficiency Virus

Transient Contacts on the Exterior of the HK97 Procapsid That Are Essential for Capsid Assembly

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