The permeability and solute transport characteristics of amphiphilic triblock-polymer vesicles containing the bacterial water-channel protein Aquaporin Z (AqpZ) were investigated. The vesicles were made of a block copolymer with symmetric poly-(2-methyloxazoline)-poly-(dimethylsiloxane)-poly-(2-methyloxazoline) (PMOXA 15-PDMS110-PMOXA 15) repeat units. Light-scattering measurements on pure polymer vesicles subject to an outwardly directed salt gradient in a stopped-flow apparatus indicated that the polymer vesicles were highly impermeable. However, a large enhancement in water productivity (permeability per unit driving force) of up to Ϸ800 times that of pure polymer was observed when AqpZ was incorporated. The activation energy (E a) of water transport for the protein-polymer vesicles (3.4 kcal/mol) corresponded to that reported for water-channel-mediated water transport in lipid membranes. The solute reflection coefficients of glucose, glycerol, salt, and urea were also calculated, and indicated that these solutes are completely rejected. The productivity of AqpZ-incorporated polymer membranes was at least an order of magnitude larger than values for existing salt-rejecting polymeric membranes. The approach followed here may lead to more productive and sustainable water treatment membranes, whereas the variable levels of permeability obtained with different concentrations of AqpZ may provide a key property for drug delivery applications.permeability ͉ triblock copolymer ͉ water treatment B iological membranes have excellent water transport characteristics, with certain membranes able to regulate permeability over a wide range. The permeability of membranes such as those present in the proximal tubules of the human kidney (1) can be increased by insertion of specific water-channel membrane proteins known as Aquaporins (AQPs). Other biological membranes, such as those in mammalian optic lenses (2), erythrocytes (3), and many other cell membranes (4) are constitutively AQP-rich. The permeabilities of AQP-rich membranes are orders of magnitude higher than those observed for unmodified phospholipid membranes (5). Additionally, some members of the AQP family have excellent solute retention capabilities for small solutes such as urea, glycerol, and glucose, even at high water transport rates (5, 6). These properties result from the unique structure of the water-selective AQPs. These AQPs have six membrane-spanning domains and a unique hourglass structure (7) with conserved charged residues that form a pore that allows both selective water transport and solute rejection. The AQP used in this study was a bacterial aquaporin from Escherichia coli, Aquaporin Z (AqpZ). AqpZ was selected because it can enhance the permeability of lipid vesicles by an order of magnitude while retaining small uncharged solutes (5). Additionally, AqpZ can be expressed in relatively large quantities in E. coli and has been reported to be quite stable under different reducing conditions and at temperatures of 4°C (5)-properties that make it att...

Surface‐immobilized nanoscale reactors utilizing membrane protein channels are used to generate precisely patterned chemically and biologically active surfaces (see image). The enzymatic conversion of a fluorogenic substrate in the cavity of immobilized nanoreactors is a model reaction demonstrating future potential application of the system in sensors, analytics, microfluidics, and single‐molecule spectroscopy.

Nanoscale devices for energy conversion require the transfer of electrons from one compartment to another. The enzyme complex I, which in vivo mediates electron transfer from NADH to ubiquinone, is an intriguing candidate for this role in nanodevices. However, complex I normally requires the presence of lipids to remain active, potentially limiting its application. Here we demonstrate for the first time that complex I can be actively reconstituted in the synthetic membrane of amphiphilic triblock copolymer vesicles. The functionality of the reconstituted protein was characterized by EPR and activity assays. Its activity is strongly influenced by the molar mass and the block length of the membrane‐forming polymers, and increases with increasing membrane thickness.magnified image

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The bioavailability limitations of proteins make them difficult to be directly delivered, particularly in diseases caused by insufficient amounts or inactive variants of those proteins. Nanoreactors represent a new promising approach to overcome these limitations because they serve both to protect the protein in their aqueous interior, and simultaneously to allow the protein to act in situ. Here we examine an antioxidant nanoreactor based on SOD encapsulated in amphiphilic block copolymer nanovesicles, and analyze its behavior as a function of the copolymer composition. The membrane of the triblock copolymer nanovesicles plays a double role, both to shield the sensitive protein and selectively to let superoxide and dioxygen penetrate to its inner space. The encapsulation efficiency for different triblock copolymer vesicles was quantified by fluorescence correlation spectroscopy using a fluorescently labeled SOD. Pulse radiolysis experiments and an enzymatic assay were used to compare the permeability of the wall-forming membranes towards superoxide anions. While the encapsulation efficiency mainly depends on the vesicle dimensions, the membrane permeability is mainly affected by the length of the hydrophobic PDMS middle blocks of our polymers. For polymers with very long PDMS chains superoxide anion transport across the membranes was too slow to be detected by our experiments.

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Ruptures of macrophage-rich atherosclerotic plaques in the coronary arteries are the main reason for heart attack. Targeted therapeutic interventions with an inhibitory effect on the macrophages promise to be beneficial, but currently available drugs such as statins achieve event reductions of only 30%. Dose-limiting adverse effects in remote organs prohibit achieving higher drug levels known to have strong inhibitory effects on macrophages. Receptor-specific targeting using statin-loaded nanometer-sized triblock copolymer vesicles with targeting moieties might allow high-dose treatment for improved efficacy, while minimizing toxicity in other cells. Vesicle uptake by target cells but not other cell types and slow intracellular content release was observed. A major improvement in biologic efficacy was observed for polymer vesicles compared to free drug, whereas no increased cytotoxicity was observed in muscle cells. Such high-dose, targeted therapy of statins through cell-specific polymer vesicles allows novel treatment paradigms not only for atherosclerosis, but appears promising for a wide range of drugs and diseases.