Enzyme encapsulation by protein cages

Chakraborti, Soumyananda; Lin, Ting‐Yu; Glatt, Sebastian; Heddle, Jonathan G.

doi:10.1039/c9ra10983h

Cited by 39 publications

(34 citation statements)

References 90 publications

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“…Many viral capsids comprise porous protein shells whose function is to protect their encapsulated genome while allowing the diffusion of water and ions across the pores 20 . The capsids of VLPs are structurally similar to the viruses from which they are derived but are non-infectious and can be repurposed to sequester a variety of different cargos including proteins, enzymes, small molecules, polymers, and even other protein compartments 21 – 25 . For example, VLPs derived from the Salmonella typhimurium virus bacteriophage P22 serve as versatile porous protein cages that can be repurposed to encapsulate a diverse array of cargo molecules.…”

Section: Introductionmentioning

confidence: 99%

Molecular exclusion limits for diffusion across a porous capsid

2021

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Molecular communication across physical barriers requires pores to connect the environments on either side and discriminate between the diffusants. Here we use porous virus-like particles (VLPs) derived from bacteriophage P22 to investigate the range of molecule sizes able to gain access to its interior. Although there are cryo-EM models of the VLP, they may not accurately depict the parameters of the molecules able to pass across the pores due to the dynamic nature of the P22 particles in the solution. After encapsulating the enzyme AdhD within the P22 VLPs, we use a redox reaction involving PAMAM dendrimer modified NADH/NAD+ to examine the size and charge limitations of molecules entering P22. Utilizing the three different accessible morphologies of the P22 particles, we determine the effective pore sizes of each and demonstrate that negatively charged substrates diffuse across more readily when compared to those that are neutral, despite the negatively charge exterior of the particles.

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Section: Introductionmentioning

confidence: 99%

Molecular exclusion limits for diffusion across a porous capsid

2021

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“…In the last two decades, protein nanocages have developed as extremely useful materials for a variety of applications including vaccine development, mostly because of their remarkable diversity in size, shape, structural biocompatibility, and immunogenicity [ 11 , 12 , 13 ]. In general, protein cages can be viewed as macromolecular containers with a wide range of cargo encapsulation and displaying abilities [ 14 , 15 , 16 ]. Among different protein-based nanocages, ferritin was the first protein cage isolated, characterized, and found very useful for a number of applications [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

“…Ferritin in general is found to be extremely stable (thermostable and protease-resistant) and biocompatible [ 19 ]. The outer and inner diameters of ferritin cages are 12 and 8 nm, respectively, and they also carry a central cavity to store iron [ 16 ]. One of the reasons ferritin is so useful for biological applications is because the surfaces of ferritin, including the inner, outer, and inter-subunit interfaces, are amenable to different types of modifications [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

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Multivalent Display of SARS-CoV-2 Spike (RBD Domain) of COVID-19 to Nanomaterial, Protein Ferritin Nanocages

et al. 2021

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SARS-CoV-2, or COVID-19, has a devastating effect on our society, both in terms of quality of life and death rates; hence, there is an urgent need for developing safe and effective therapeutics against SARS-CoV-2. The most promising strategy to fight against this deadly virus is to develop an effective vaccine. Internalization of SARS-CoV-2 into the human host cell mainly occurs through the binding of the coronavirus spike protein (a trimeric surface glycoprotein) to the human angiotensin-converting enzyme 2 (ACE2) receptor. The spike-ACE2 protein–protein interaction is mediated through the receptor-binding domain (RBD) of the spike protein. Mutations in the spike RBD can significantly alter interactions with the ACE2 host receptor. Due to its important role in virus transmission, the spike RBD is considered to be one of the key molecular targets for vaccine development. In this study, a spike RBD-based subunit vaccine was designed by utilizing a ferritin protein nanocage as a scaffold. Several fusion protein constructs were designed in silico by connecting the spike RBD via a synthetic linker (different sizes) to different ferritin subunits (H-ferritin and L-ferritin). The stability and the dynamics of the engineered nanocage constructs were tested by extensive molecular dynamics simulation (MDS). Based on our MDS analysis, a five amino acid-based short linker (S-Linker) was the most effective for displaying the spike RBD over the surface of ferritin. The behavior of the spike RBD binding regions from the designed chimeric nanocages with the ACE2 receptor was highlighted. These data propose an effective multivalent synthetic nanocage, which might form the basis for new vaccine therapeutics designed against viruses such as SARS-CoV-2.

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“…8,9 In recent years both natural and artificial protein cages have been developed for numerous potential applications including in biotechnology and as therapeutics. [10][11][12] Among cages that have been developed in this way, ferritin has been perhaps the most extensively used including for cargo encapsulation, metal mineralisation, bio-sensors, and targeted drug delivery. [13][14][15] Ferritin is an iron storage protein found near ubiquitously across all forms of life.…”

Section: Introductionmentioning

confidence: 99%