Tuning the catalytic properties of P22 nanoreactors through compositional control

Sharma, Jhanvi; Douglas, Trevor

doi:10.1039/c9nr08348k

Cited by 43 publications

(78 citation statements)

References 91 publications

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“…The focus of P22 research has not only been on cargo encapsulation, but the dynamics and stability of the assembled capsids [210,211,217,218] as well as in vitro cargo retention, release and incorporation [219–221] have been extensively studied as well. Interestingly, the isolated P22 capsid can undergo several rearrangements leading to cargo release upon temperature triggers [210,211] .…”

Section: Artificial Organelles Produced In Vivomentioning

confidence: 99%

“…Treatment with sodium dodecyl sulfate [217] or guanidine hydrochloride [219] can trigger the release from SP as well. Via these approaches the packaging density of the P22 capsids can be varied and the catalytic properties of an alcohol dehydrogenase (AdhD) have been controlled [221] . Although the demonstrated control over P22 dynamics and interior density can be quite easily translated into control over catalytic activity under in vitro conditions, the requirement for high temperatures and high chemical concentrations does not quite work for in vivo studies.…”

Section: Artificial Organelles Produced In Vivomentioning

confidence: 99%

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Artificial Organelles: Towards Adding or Restoring Intracellular Activity

2021

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Compartmentalization is one of the main characteristics that define living systems. Creating a physically separated microenvironment allows nature a better control over biological processes, as is clearly specified by the role of organelles in living cells. Inspired by this phenomenon, researchers have developed a range of different approaches to create artificial organelles: compartments with catalytic activity that add new function to living cells. In this review we will discuss three complementary lines of investigation. First, orthogonal chemistry approaches are discussed, which are based on the incorporation of catalytically active transition metal‐containing nanoparticles in living cells. The second approach involves the use of premade hybrid nanoreactors, which show transient function when taken up by living cells. The third approach utilizes mostly genetic engineering methods to create bio‐based structures that can be ultimately integrated with the cell's genome to make them constitutively active. The current state of the art and the scope and limitations of the field will be highlighted with selected examples from the three approaches.

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Section: Artificial Organelles Produced In Vivomentioning

confidence: 99%

Section: Artificial Organelles Produced In Vivomentioning

confidence: 99%

Artificial Organelles: Towards Adding or Restoring Intracellular Activity

2021

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“…The catalytic activity of encapsulated ADH was finely tuned by controlling the stoichiometry of enzyme loading and packing density of enzymes inside the P22 VLPs [ 90 ]. Compositional control was achieved by mixing different ratios of ADH fused SP (ADH-SP) and wild type SPs (wtSPs) during in vitro assembly of P22 VLPs whereas packing density was controlled by selective removal of wtSPs through P22 capsid pores using mild treatment with a chaotrope.…”

Section: Protein Scaffolds Employed In Multi-enzyme Assemblymentioning

confidence: 99%

“…Compositional control was achieved by mixing different ratios of ADH fused SP (ADH-SP) and wild type SPs (wtSPs) during in vitro assembly of P22 VLPs whereas packing density was controlled by selective removal of wtSPs through P22 capsid pores using mild treatment with a chaotrope. The wtSPs were observed to exert a crowding effect on the enzyme which finely tuned the enzymatic output [ 90 ]. A similar approach was used to synthesize complex proteinaceous hierarchical structures with P22 VLPs that mimic cellular environments having different macromolecules and subcellular compartments [ 91 ].…”

Section: Protein Scaffolds Employed In Multi-enzyme Assemblymentioning

confidence: 99%

Protein scaffolds: A tool for multi-enzyme assembly

Gad

Ayakar

2021

Biotechnology Reports

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Highlights Protein scaffold acts as a backbone with several interacting domains and ligands for the integration of specific enzymes. Different protein-enzyme ligation strategies are used for the co-localization of enzymes. Scaffolds permit control over spatial organization, enzyme stoichiometry, and proximity. Improvement in pathway flux and product yield can be achieved with scaffold-based multi-enzyme complex. Various analytical tools are utilized to study the multi-enzyme assembly process.

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“…Furthermore, the modular nature of the in vitro approach allows encapsulation of a cargo whose folding may go awry prior to encapsulation and requires external refolding to exhibit desired activity [27]. This approach can now be extended to a variety of other cargos including cages, such as ferritin, to create hierarchical multi-compartment structures [34,35]. Encapsulation of StAv generates a non-covalent conjugation site to which biotin-tagged molecules are easily attached for interior or exterior loading.…”

Section: Synthesis Of P22-stav Vlpmentioning

confidence: 99%

Nano-Particulate Platforms for Vaccine Delivery to Enhance Antigen-Specific CD8+ T-Cell Response

et al. 2021

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Vaccines remain the most effective way to protect populations against deathly infectious diseases. Several disadvantages associated with the traditional vaccines that use whole pathogens have led to the development of alternative strategies including the use of recombinant subunit vaccines. Subunit vaccines are, in general, safer than whole pathogens but tend to be less immunogenic due to the lack of molecular cues that are typically found on whole pathogens. To enhance immunogenicity, the subunit antigen can be administered with adjuvants that stimulate the innate immune system as a means to steer the quality and magnitude of the adaptive immune response. Novel classes of adjuvants are formulated using particle-based platforms such as virus-like particles, liposomes, and polymeric nanoparticles. These particle-based systems present antigens in ways reminiscent of whole pathogens. Such platforms offer several advantages that include co-delivery of antigen along with innate immune stimulators in a highly immunogenic format. Here we describe our recent efforts to synthesize, characterize, and validate two promising nanoparticle-based delivery systems and demonstrate their potential to induce antigen-specific CD8 + T cell responses, essential in clearing infection with intracellular pathogens, such as viruses and bacteria, and eradicating tumors.

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Tuning the catalytic properties of P22 nanoreactors through compositional control

Abstract: Fine-tuning of the catalytic activity of enzyme based P22 nanoreactors through compositional control.

Cited by 43 publications

References 91 publications

Artificial Organelles: Towards Adding or Restoring Intracellular Activity

Artificial Organelles: Towards Adding or Restoring Intracellular Activity

Protein scaffolds: A tool for multi-enzyme assembly

Nano-Particulate Platforms for Vaccine Delivery to Enhance Antigen-Specific CD8+ T-Cell Response

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