skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Virus-Like Particles (VLPs) as a Platform for Hierarchical Compartmentalization

Journal Article · · Biomacromolecules

Hierarchically self-assembled structures are common in biology, but it is often challenging to design and fabricate synthetic analogs. The archetypal cell is defined by hierarchically organized multicompartmentalized structures with boundaries that delineate the interior from exterior environments and is an inspiration for complex functional materials. Here, we have demonstrated an approach to the design and construction of a nested protein cage system that can additionally incorporate the packing of other functional macromolecules and exhibit some of the features of a minimal synthetic cell-like material. We have demonstrated a strategy for controlled co-packaging of subcompartments, ferritin (Fn) cages, together with active cellobiose-hydrolyzing β-glycosidase enzyme macromolecules, CelB, inside the sequestered volume of the bacteriophage P22 capsid. Using controlled in vitro assembly, we were able to modulate the stoichiometry of Fn cages and CelB encapsulated inside the P22 to control the degree of compartmentalization. The co-encapsulated enzyme CelB showed catalytic activity even when packaged at high total macromolecular concentrations comparable to an intracellular environment. Here, this approach could be used as a model to create synthetic protein-based protocells that can confine smaller functionalized proto-organelles and additional macromolecules to support a range of biochemical reactions.

Research Organization:
Indiana Univ., Bloomington, IN (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences & Engineering Division; National Science Foundation (NSF)
Grant/Contract Number:
SC0016155; NSF-BMAT DMR-1507282; NSF-NM CMMI-1922883
OSTI ID:
1716721
Journal Information:
Biomacromolecules, Vol. 21, Issue 6; ISSN 1525-7797
Publisher:
American Chemical SocietyCopyright Statement
Country of Publication:
United States
Language:
English

References (82)

Engineering Bacterial Microcompartment Shells: Chimeric Shell Proteins and Chimeric Carboxysome Shells journal July 2014
Designs for life: protocell models in the laboratory journal January 2012
Synthetic cellularity based on non-lipid micro-compartments and protocell models journal October 2014
A positive definition of prokaryotes journal August 2006
Bacterial microcompartments and the modular construction of microbial metabolism journal January 2015
Self-assembly and optically triggered disassembly of hierarchical dendron–virus complexes journal March 2010
A Metal Organic Framework with Spherical Protein Nodes: Rational Chemical Design of 3D Protein Crystals journal September 2015
Binary Protein Crystals for the Assembly of Inorganic Nanoparticle Superlattices journal September 2016
Higher Order Assembly of Virus-like Particles (VLPs) Mediated by Multi-valent Protein Linkers journal January 2015
Modular Self-Assembly of Protein Cage Lattices for Multistep Catalysis journal November 2017
Encapsulation of an Enzyme Cascade within the Bacteriophage P22 Virus-Like Particle journal November 2013
Bottom-up Construction of a Primordial Carboxysome Mimic journal August 2016
Assembling Enzymatic Cascade Pathways inside Virus-Based Nanocages Using Dual-Tasking Nucleic Acid Tags journal January 2017
Prokaryotic nanocompartments form synthetic organelles in a eukaryote journal April 2018
Construction of Matryoshka-Type Structures from Supercharged Protein Nanocages journal November 2014
Droplet-based microfluidics for artificial cell generation: a brief review journal August 2016
Construction of membrane-bound artificial cells using microfluidics: a new frontier in bottom-up synthetic biology journal June 2016
Microfluidic generation of encapsulated droplet interface bilayer networks (multisomes) and their use as cell-like reactors journal January 2016
Artificial cell mimics as simplified models for the study of cell biology journal June 2017
Stepwise Synthesis of Giant Unilamellar Vesicles on a Microfluidic Assembly Line journal March 2011
Layer-by-layer cell membrane assembly journal September 2013
In vitro gene expression and enzyme catalysis in bio-inorganic protocells journal January 2011
Interfacial assembly of protein–polymer nano-conjugates into stimulus-responsive biomimetic protocells journal July 2013
Fatty acid membrane assembly on coacervate microdroplets as a step towards a hybrid protocell model journal April 2014
Polymersomes in “Gelly” Polymersomes: Toward Structural Cell Mimicry journal December 2011
Polymersomes in Polymersomes: Multiple Loading and Permeability Control journal December 2011
Polymer Vesicles journal August 2002
Mimicking Cellular Compartmentalization in a Hierarchical Protocell through Spontaneous Spatial Organization journal July 2019
Encapsulation of bilayer vesicles by self-assembly journal May 1997
The Vesosome - A Multicompartment Drug Delivery Vehicle journal January 2004
Multiple Lipid Compartments Slow Vesicle Contents Release in Lipases and Serum journal September 2007
Fabrication of PLG microspheres with precisely controlled and monodisperse size distributions journal May 2001
Uniform double-walled polymer microspheres of controllable shell thickness journal April 2004
Why and how to prepare biodegradable, monodispersed, polymeric microparticles in the field of pharmacy? journal April 2011
One-Pot Assembly of Complex Giant Unilamellar Vesicle-Based Synthetic Cells journal April 2019
Photosynthetic artificial organelles sustain and control ATP-dependent reactions in a protocellular system journal May 2018
Multicompartmentalized polymersomes for selective encapsulation of biomacromolecules journal January 2011
Remote Control of Bioreactions in Multicompartment Capsules journal October 2007
Shell-in-Shell Microcapsules: A Novel Tool for Integrated, Spatially Confined Enzymatic Reactions journal July 2007
Production of uniform droplets using membrane, microchannel and microfluidic emulsification devices journal February 2012
Particle control of emulsion by membrane emulsification and its applications journal December 2000
Multicompartment Particle Assemblies for Bioinspired Encapsulated Reactions journal October 2011
Structural Studies of the Sputnik Virophage journal November 2009
Structural Studies of the Giant Mimivirus journal April 2009
Structural basis of enzyme encapsulation into a bacterial nanocompartment journal August 2008
Assembly in vitro of Rhodococcus jostii  RHA1 encapsulin and peroxidase DypB to form a nanocompartment journal April 2013
A virus capsid‐like nanocompartment that stores iron and protects bacteria from oxidative stress journal July 2014
Comparison of the three-dimensional structures of recombinant human H and horse L ferritins at high resolution 1 1Edited by R. Huber journal May 1997
The ferritin superfamily: Supramolecular templates for materials synthesis journal August 2010
Enzyme Encapsulation by a Ferritin Cage journal October 2017
Virus-like particle nanoreactors: programmed encapsulation of the thermostable CelB glycosidase inside the P22 capsid journal January 2012
Developing a Dissociative Nanocontainer for Peptide Drug Delivery journal October 2015
Atom transfer radical polymerization on the interior of the P22 capsid and incorporation of photocatalytic monomer crosslinks journal October 2013
Co-localization of catalysts within a protein cage leads to efficient photochemical NADH and/or hydrogen production journal January 2016
Mechanism of head assembly and DNA encapsulation in Salmonella phage P22 journal November 1973
Accurate model annotation of a near-atomic resolution cryo-EM map journal March 2017
Genetically Programmed In Vivo Packaging of Protein Cargo and Its Controlled Release from Bacteriophage P22 journal June 2011
Modular interior loading and exterior decoration of a virus-like particle journal January 2017
Estimation of macromolecule concentrations and excluded volume effects for the cytoplasm of Escherichia coli journal December 1991
Targeting of Cancer Cells with Ferrimagnetic Ferritin Cage Nanoparticles journal December 2006
A human ferritin iron oxide nano-composite magnetic resonance contrast agent journal November 2008
Penton Release from P22 Heat-Expanded Capsids Suggests Importance of Stabilizing Penton-Hexon Interactions during Capsid Maturation journal April 2003
Scaffolding protein regulates the polymerization of P22 coat subunits into icosahedral shells in vitro journal August 1988
Implementation of P22 Viral Capsids as Nanoplatforms journal October 2010
Folding of the phage P22 coat protein in vitro journal October 1993
Purification of the coat and scaffolding proteins from procapsids of bacteriophage P22 journal July 1981
Fiji: an open-source platform for biological-image analysis journal June 2012
Automated molecular microscopy: The new Leginon system journal July 2005
Fully automated, sequential tilt-series acquisition with Leginon journal July 2009
Computer Visualization of Three-Dimensional Image Data Using IMOD journal January 1996
UCSF Chimera?A visualization system for exploratory research and analysis journal January 2004
Self-assembling biomolecular catalysts for hydrogen production journal December 2015
Design of a VLP-nanovehicle for CYP450 enzymatic activity delivery journal October 2015
Cargo–shell and cargo–cargo couplings govern the mechanics of artificially loaded virus-derived cages journal January 2016
Nanoreactors by Programmed Enzyme Encapsulation Inside the Capsid of the Bacteriophage P22 journal May 2012
Core-controlled polymorphism in virus-like particles journal January 2007
Cargo Retention inside P22 Virus-Like Particles journal August 2018
Mechanism of ferritin iron uptake: activity of the H-chain and deletion mapping of the ferro-oxidase site. A study of iron uptake and ferro-oxidase activity of human liver, recombinant H-chain ferritins, and of two H-chain deletion mutants. journal December 1988
Templated assembly of organic–inorganic materials using the core shell structure of the P22 bacteriophage journal January 2011
Chemically Induced Morphogenesis of P22 Virus-like Particles by the Surfactant Sodium Dodecyl Sulfate journal November 2018
Tuning the catalytic properties of P22 nanoreactors through compositional control journal January 2020
Join the crowd journal September 2003