Stimuli-Triggered Activity of Nanoreactors by Biomimetic Engineering Polymer Membranes

Einfalt, Tomaz; Goers, Roland; Dinu, Ionel Adrian; Najer, Adrian; Spulber, Mariana; Onaca-Fischer, Ozana; Palivan, Cornelia G.

doi:10.1021/acs.nanolett.5b03386

Cited by 82 publications

(96 citation statements)

References 47 publications

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“…As FCS only detects objects associated with PR-GFP, we could potentially detect different vesicle populations. Evidence for a good correlation of sizes obtained by DLS/FCS and micrographs haven been presented for liposomes 36,37 and PDMS-PMOXA polymersomes [38][39][40][41] , even though direct methods like (cryo-)TEM are considered more precise. The polydispersity index (PdI), obtained from cumulants analysis, was utilized as a measure for homogeneity.…”

Section: Resultsmentioning

confidence: 99%

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Optimized reconstitution of membrane proteins into synthetic membranes

et al. 2018

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Light-driven proton pumps, such as proteorhodopsin, have been proposed as an energy source in the field of synthetic biology. Energy is required to power biochemical reactions within artificially created reaction compartments like proto-or nanocells, which are typically based on either lipid or polymer membranes. The insertion of membrane proteins into these membranes is delicate and quantitative studies comparing these two systems are needed. Here we present a detailed analysis of the formation of proteoliposomes and proteopolymersomes and the requirements for a successful reconstitution of the membrane protein proteorhodopsin. To this end, we apply design of experiments to provide a mathematical framework for the reconstitution process. Mathematical optimization identifies suitable reconstitution conditions for lipid and polymer membranes and the obtained data fits well to the predictions. Altogether, our approach provides experimental and modeling evidence for different reconstitution mechanisms depending on the membrane type which resulted in a surprisingly similar performance.

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Section: Resultsmentioning

confidence: 99%

“…The FCS measurements were performed as already described 40 . Briefly, an inverted microscope (Axiovert 200 M, Zeiss), equipped with a laser scanning microscopy module LSM 510 (Zeiss) and a ConfoCor2 (Zeiss) module was used.…”

Section: Methodsmentioning

confidence: 99%

Optimized reconstitution of membrane proteins into synthetic membranes

et al. 2018

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“…proteolytic attack), and allow them to act 'in situ'. Polymer nanoreactors based on a single enzyme type have been developed with various proteins, such as horseradish peroxidase, [11,20] nucleoside hydrolase, [23] or acid phosphatase. [24] Interestingly, by encapsulating enzymes with dual-functions, such as haemoglobin, it was possible to design multifunctional nanoreactors to simultaneously transport oxygen and degrade peroxynitrites.…”

Section: Bioinspired Permeabilization Of Polymer Membranesmentioning

confidence: 99%

“…4). [19,20] Such nanoreactors with triggered activity represent ideal candidates when a specific step in the flow of a molecular factory requires a reaction activated 'on demand' by the presence of a stimulus.…”

Section: Introductionmentioning

confidence: 99%

Artificial Organelles: Reactions inside Protein–Polymer Supramolecular Assemblies

Garni

Einfalt

Lomora

et al. 2016

Chimia

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Reactions inside confined compartments at the nanoscale represent an essential step in the development of complex multifunctional systems to serve as molecular factories. In this respect, the biomimetic approach of combining biomolecules (proteins, enzymes, mimics) with synthetic membranes is an elegant way to create functional nanoreactors, or even simple artificial organelles, that function inside cells after uptake. Functionality is provided by the specificity of the biomolecule(s), whilst the synthetic compartment provides mechanical stability and robustness. The availability of a large variety of biomolecules and synthetic membranes allows the properties and functionality of these reaction spaces to be tailored and adjusted for building complex self-organized systems as the basis for molecular factories.

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“…ii) What are the optimal concentrations could traverse the ∼3.3 nm-diameter outer membrane protein F (OmpF) to diffuse into the polymersomes. [13b] To further biocatalyze reactions, [21] OmpF was chemically modified with a molecular 'cap' that acted as a pH responsive gate and could be opened by lowering the pH. In this manner, a modified OmpF-bearing polymersome could serve as a nanoreactor by converting the influx of chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) using a pre-encapsulated model enzyme horseradish peroxidase (HRP) within its lumen.…”

Section: Nanoreactors Artificial Organelles and Molecular Factoriesmentioning

confidence: 99%

Nucleocytoplasmic Transport: A Paradigm for Molecular Logistics in Artificial Systems

Vujica¹,

Zelmer²,

Panatala³

et al. 2016

Chimia

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Artificial organelles, molecular factories and nanoreactors are membrane-bound systems envisaged to exhibit cell-like functionality. These constitute liposomes, polymersomes or hybrid lipo-polymersomes that display different membrane-spanning channels and/or enclose molecular modules. To achieve more complex functionality, an artificial organelle should ideally sustain a continuous influx of essential macromolecular modules (i.e. cargoes) and metabolites against an outflow of reaction products. This would benefit from the incorporation of selective nanopores as well as specific trafficking factors that facilitate cargo selectivity, translocation efficiency, and directionality. To wards this goal, we describe how proteinaceous cargoes are transported between the nucleus and cytoplasm by nuclear pore complexes and the biological trafficking machinery in living cells (i.e. nucleocytoplasmic transport). On this basis, we discuss how biomimetic control may be implemented to selectively import, compartmentalize and accumulate diverse macromolecular modules against concentration gradients in artificial organelles.

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Stimuli-Triggered Activity of Nanoreactors by Biomimetic Engineering Polymer Membranes

Cited by 82 publications

References 47 publications

Optimized reconstitution of membrane proteins into synthetic membranes

Optimized reconstitution of membrane proteins into synthetic membranes

Artificial Organelles: Reactions inside Protein–Polymer Supramolecular Assemblies

Nucleocytoplasmic Transport: A Paradigm for Molecular Logistics in Artificial Systems

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