Electrostatic assembly of a multicomponent peptide/amphiphile nanotube.

The ability to integrate the elements of a multicomponent nanostructure with nanoscale precision by co-assembly provides a versatile strategy to create novel materials with tunable properties. The search for function in these materials will require new strategies to be developed that control the assembly process, especially for structurally dissimilar components, which often have a propensity to self-sort into non-integrated nanostructures. In this work, two components, a peptide (1) and an amphiphile (2), were integratively co-assembled into a multicomponent nanotube. The interaction between the two components at the supramolecular level was driven by the electrostatic complementarity of the components, which was controlled by the pH-dependent charge of 1. Characterization of the co-assembled nanotube, 1-2NT, was achieved using a combination of TEM, AFM, CLSM and SIM techniques, which showed that both components were colocalized within the nanotube. These studies, in conjunction with CD, IR and fluorescence studies, suggested that 1 and 2 were arranged in partially reorganized, self-sorted domains, which were integrated as laminated nanoribbons that coiled together into the final co-assembled nanotube.

Dissipative self-assembly of a proline catalyst for temporal regulation of the aldol reaction.

The spatiotemporal regulation of chemical reactivity in biological systems permits a network of metabolic reactions to take place within the same cellular environment. The exquisite control of reactivity is often mediated by out-of-equilibrium structures that remain functional only as long as fuel is present to maintain the higher energy, active state. An important goal in supramolecular chemistry aims to develop functional, energy dissipating systems that approach the sophistication of biological machinery. The challenge is to create strategies that couple the energy consumption needed to promote a molecule to a higher energy, assembled state to a functional property such as catalytic activity. In this work, we demonstrated that the assembly of a spiropyran (SP) dipeptide (1) transiently promoted the proline-catalyzed aldol reaction in water when visible light was present as fuel. The transient catalytic activity emerged from 1 under light illumination due to the photoisomerization of the monomeric, O-protonated (1-MCH+) merocyanine form to the spiropyran (1-SP) state, which rapidly assembled into nanosheets capable of catalyzing the aldol reaction in water. When the light source was removed, thermal isomerization to the more stable MCH+ form caused the nanosheets to dissociate into a catalytically inactive, monomeric state. Under these conditions, the aldol reaction could be repeatedly activated and deactivated by switching the light source on and off.

Self-assembly of a 5-fluorouracil and camptothecin dual drug dipeptide conjugate.

Nano-formulated, combinatory therapeutics that control the spatiotemporal aspects of drug release have potential to overcome many of the challenges faced in cancer therapy. Herein, we describe a peptide nanotube functionalized with two anticancer drugs, 5-fluoruracil (5-FU) and camptothecin (CPT). The nanotube was formed via peptide self-assembly, which positioned 5-FU on the surface at the aqueous interface; whereas, CPT was sequestered within the hydrophobic walls. Thus, two different release profiles were observed: rapid release of 5-FU, followed by slower, sustained production of CPT. This profile emerged from the rapid hydrolytic cleavage of 5-FU at the aqueous/nanotube interface, which produced a smaller nanotube comprised of the peptide fragment.

Co-assembly of a multicomponent network of nanofiber-wrapped nanotubes.

Strategies to create organized multicomponent nanostructures composed of discrete, self-sorted domains are important for developing materials that mimic the complexity and multifunctionality found in biological systems. These structures can be challenging to achieve due to the required balance of molecular self-recognition and supramolecular attraction needed between the components. Herein, we report a strategy to construct a two-component nanostructure via a hierarchical assembly process whereby two monomeric building blocks undergo self-sorting assembly at the molecular level followed by a supramolecular association to form a nanofiber-wrapped nanotube. The two molecules self-sorted into respective nanofiber and nanotube assemblies, yet assembly of the nanofibers in the presence of the nanotube template allowed for directed integration into a hierarchical multilayer structure via electrostatic interactions. The fiber-wrapped nanotube co-assembly was characterized using transmission electron microscopy (TEM), atomic force microscopy (AFM) and Förster resonance energy transfer (FRET) between the components. Strategies to co-assemble multicomponent nanostructures composed of discrete, spatially sorted domains with controllable higher level interactions will be critical for the development of novel, functionally competent nanomaterials.

Light-driven dissipative self-assembly of a peptide hydrogel.

Light energy provides an attractive fuel source for energy dissipating systems because of the lack of waste production, wavelength tunability and the potential for spatial and temporal resolution. In this work, we describe a peptide-spiropyran conjugate that assembled into a transient nanofiber hydrogel in the presence of visible light, and dissociated when the light source was removed.

Amino Acid Nanofibers Improve Glycemia and Confer Cognitive Therapeutic Efficacy to Bound Insulin.

Diabetes poses a high risk for debilitating complications in neural tissues, regulating glucose uptake through insulin-dependent and predominantly insulin-independent pathways. Supramolecular nanostructures provide a flexible strategy for combinatorial regulation of glycemia. Here, we compare the effects of free insulin to insulin bound to positively charged nanofibers comprised of self-assembling amino acid compounds (AACs) with an antioxidant-modified side chain moiety (AAC2) in both in vitro and in vivo models of type 1 diabetes. Free AAC2, free human insulin (hINS) and AAC2-bound-human insulin (AAC2-hINS) were tested in streptozotocin (STZ)-induced mouse model of type 1 diabetes. AAC2-hINS acted as a complex and exhibited different properties compared to free AAC2 or hINS. Mice treated with the AAC2-hINS complex were devoid of hypoglycemic episodes, had improved levels of insulin in circulation and in the brain, and increased expression of neurotransmitter taurine transporter, Slc6a6. Consequently, treatment with AAC2-hINS markedly advanced both physical and cognitive performance in mice with STZ-induced and genetic type 1 diabetes compared to treatments with free AAC2 or hINS. This study demonstrates that the flexible nanofiber AAC2 can serve as a therapeutic platform for the combinatorial treatment of diabetes and its complications.

The impact of metal coordination on the assembly of bis(indolyl)methane-naphthalene-diimide amphiphiles.

The self-assembly and coordination of amphiphiles comprised of naphthalenediimide (NDI) and bis(indolyl)methane (BIM) chromophores were investigated as a function of pH and metal. As observed by TEM, SEM and AFM imaging, the self-assembly of NDI-BIM 1 produced irregular nanostructures at neutral pH in CH3CN-H2O (1 : 1); whereas, well-defined nanotubes were observed at pH 2. Conversely, Fmoc-protected, NDI-BIM 2 formed nanotubes at neutral pH and nonspecific aggregates at pH 2. Upon coordination of Cu2+ ions to the bis(indoyl)methane moiety, a reorganization from nanotubes to vesicular structures was observed.

Self-assembly of a robust, reduction-sensitive camptothecin nanotube.

The self-assembly and covalent crosslinking of a camptothecin (CPT) tetrapeptide nanotube is reported. Intermolecular disulfide bond formation of a self-assembled CPT-peptide reversibly stabilized the nanotubes toward dissociation at low concentrations, resulting in inhibited release of CPT. In the presence of dithiothreitol (DTT), the release of CPT was significantly accelerated. The crosslinked nanotubes also exhibited in vitro cytotoxicity against human non-small cell lung cancer cell lines A549 and H460.

Light-controlled self-assembly of a dithienylethene bolaamphiphile in water.

The self-assembly of bolaamphiphiles comprised of a central photochromic dithienylethene (DTE) chromophore was investigated in aqueous media. Irradiation at 254 nm induced a conversion from the open to closed states of the DTE chromophores. Whereas, in water, irradiation produced a photostationary state of 20 : 80 (open/closed), in methanol the ratio improved to 10 : 90 (open/closed). The open → closed transition was accompanied by the formation of 1D nanofibers during incubation in darkness.

Enhanced Stability of Peptide Nanofibers Coated with a Conformal Layer of Polydopamine.

The susceptibility of self-assembled materials to changes of environmental conditions and mechanical forces often limits their utility for many applications. In this work, the surface of nanofibers formed by ß-sheet peptide self-assemblies were coated by polydopamine (PDA) deposition. This conformal coating process rendered the nanofiber dimensions and internal π-stacking chirality impervious to changes in pH, temperature, and physical processing by spin-coating onto a silicon wafer. Whereas sonication-induced shearing of the dopamine/naphthalenediimide-dilysine (DA/NDI-KK) composite irreversibly shortened the nanofibers into 100-200 nm segments, the uncoated nanofibers unraveled into single strands upon similar treatment. Additionally, the PDA-coated nanofibers could be wrapped by an additional layer comprised of a positively charged polyelectrolyte polymer.

Amino acid-based compound activates atypical PKC and leptin receptor pathways to improve glycemia and anxiety like behavior in diabetic mice.

Differences in glucose uptake in peripheral and neural tissues account for the reduced efficacy of insulin in nervous tissues. Herein, we report the design of short peptides, referred as amino acid compounds (AAC) with and without a modified side chain moiety. At nanomolar concentrations, a candidate therapeutic molecule, AAC2, containing a 7-(diethylamino) coumarin-3-carboxamide side-chain improved glucose control in human peripheral adipocytes and the endothelial brain barrier cells by activation of insulin-insensitive glucose transporter 1 (GLUT1). AAC2 interacted specifically with the leptin receptor (LepR) and activated atypical protein kinase C zeta (PKCς) to increase glucose uptake. The effects induced by AAC2 were absent in leptin receptor-deficient predipocytes and in Leprdb mice. In contrast, AAC2 established glycemic control altering food intake in leptin-deficient Lepob mice. Therefore, AAC2 activated the LepR and acted in a cytokine-like manner distinct from leptin. In a monogenic Ins2Akita mouse model for the phenotypes associated with type 1 diabetes, AAC2 rescued systemic glucose uptake in these mice without an increase in insulin levels and adiposity, as seen in insulin-treated Ins2Akita mice. In contrast to insulin, AAC2 treatment increased brain mass and reduced anxiety-related behavior in Ins2Akita mice. Our data suggests that the unique mechanism of action for AAC2, activating LepR/PKCς/GLUT1 axis, offers an effective strategy to broaden glycemic control for the prevention of diabetic complications of the nervous system and, possibly, other insulin insensitive or resistant tissues.

Threading carbon nanotubes through a self-assembled nanotube.

Achieving the co-assembly of more than one component represents an important challenge in the drive to create functional self-assembled nanomaterials. Multicomponent nanomaterials comprised of several discrete, spatially sorted domains of components with high degrees of internal order are particularly important for applications such as optoelectronics. In this work, single-walled carbon nanotubes (SWNTs) were threaded through the inner channel of nanotubes formed by the bolaamphiphilic self-assembly of a naphthalenediimide-lysine (NDI-Bola) monomer. The self-assembly process was driven by electrostatic interactions, as indicated by ζ-potential measurements, and cation-π interactions between the surface of the SWNT and the positively charged, NDI-Bola nanotube interior. To increase the threading efficiency, the NDI-Bola nanotubes were fragmented into shortened segments with lengths of <100 nm via sonication-induced shear, prior to co-assembly with the SWNTs. The threading process created an initial composite nanostructure in which the SWNTs were threaded by multiple, shortened segments of the NDI-Bola nanotube that progressively re-elongated along the SWNT surface into a continuous radial coating around the SWNT. The resultant composite structure displayed NDI-Bola wall thicknesses twice that of the parent nanotube, reflecting a bilayer wall structure, as compared to the monolayer structure of the parent NDI-Bola nanotube. As a final, co-axial outer layer, poly(p-phenyleneethynylene) (PPE-SO3Na, M W = 5.76 × 104, PDI - 1.11) was wrapped around the SWNT/NDI-Bola composite resulting in a three-component (SWNT/NDI-Bola/PPE-SO3Na) composite nanostructure.

pH-Controlled Chiral Packing and Self-Assembly of a Coumarin Tetrapeptide.

A coumarin-tetrapeptide conjugate, EFEK(DAC)-NH2 (1), is reported to undergo a pH-dependent interconversion between nanotubes and nanoribbons. An examination of zeta potential measurements, circular dichroism (CD) spectra, and microscopy imaging (transmission electron microscopy and atomic force microscopy) identified three different self-assembly regimes based on pH: (1) pH 2-5, positively charged, left-handed helical nanotubes; (2) pH 6-8, negatively charged, right-handed helical nanoribbons; and (3) pH ≥ 9.0, a monomeric/disassembled peptide. The nanotubes exhibited uniform diameters of 41 ± 5 nm and wall thicknesses of 4.8 ± 0.8 nm, whereas the nanoribbons existed as either flat or twisted sheets ranging in width from 11 to 60 nm with heights of 8 ± 1 nm. The UV-vis and CD spectra of the most common antiparallel, ß-sheet conformation of 1-dimer were simulated at the B3LYP/def2svpd level of theory in implicit water. These studies indicated that the transition from nanotubes to nanoribbons was coupled to an M → P helical inversion of the coumarin packing orientation, respectively, within the nanostructures. The assembly process was driven by ß-sheet aggregation and π-π interactions, leading to the formation of nanoribbons, which progressively wound into helical ribbons and laterally grew into smooth nanotubes as the pH decreased.

Controlling the length of self-assembled nanotubes by sonication followed by polymer wrapping.

This work demonstrates that sonication, followed by polymer-wrapping, is an effective strategy to modulate the length of self-assembled nanotubes. The length distributions of the nanotubes were controlled by varying the amplitude of sonication. Wrapping the nanotubes with ionic polymers suspended the propensity of the nanotube fragments to re-assemble over time into their elongated precursors.

Strategy for the Co-Assembly of Co-Axial Nanotube-Polymer Hybrids.

Nanostructured materials having multiple, discrete domains of sorted components are particularly important to create efficient optoelectronics. The construction of multicomponent nanostructures from self-assembled components is exceptionally challenging due to the propensity of noncovalent materials to undergo structural reorganization in the presence of excipient polymers. This work demonstrates that polymer-nanotube composites comprised of a self-assembled nanotube wrapped with two conjugated polymers could be assembled using a layer-by-layer approach. The polymer-nanotube nanostructures arrange polymer layers coaxially on the nanotube surface. Femtosecond transient absorption (TA) studies indicated that the polymer-nanotube composites undergo photoinduced charge separation upon excitation of the NDI chromophore within the nanotube.

Synthetic CO2-fixation enzyme cascades immobilized on self-assembled nanostructures that enhance CO2/O2 selectivity of RubisCO.

BACKGROUND: With increasing concerns over global warming and depletion of fossil-fuel reserves, it is attractive to develop innovative strategies to assimilate CO2, a greenhouse gas, into usable organic carbon. Cell-free systems can be designed to operate as catalytic platforms with enzymes that offer exceptional selectivity and efficiency, without the need to support ancillary reactions of metabolic pathways operating in intact cells. Such systems are yet to be exploited for applications involving CO2 utilization and subsequent conversion to valuable products, including biofuels. The Calvin-Benson-Bassham (CBB) cycle and the enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) play a pivotal role in global CO2 fixation. RESULTS: We hereby demonstrate the co-assembly of two RubisCO-associated multienzyme cascades with self-assembled synthetic amphiphilic peptide nanostructures. The immobilized enzyme cascades sequentially convert either ribose-5-phosphate (R-5-P) or glucose, a simpler substrate, to ribulose 1,5-bisphosphate (RuBP), the acceptor for incoming CO2 in the carboxylation reaction catalyzed by RubisCO. Protection from proteolytic degradation was observed in nanostructures associated with the small dimeric form of RubisCO and ancillary enzymes. Furthermore, nanostructures associated with a larger variant of RubisCO resulted in a significant enhancement of the enzyme's selectivity towards CO2, without adversely affecting the catalytic activity. CONCLUSIONS: The ability to assemble a cascade of enzymes for CO2 capture using self-assembling nanostructure scaffolds with functional enhancements show promise for potentially engineering entire pathways (with RubisCO or other CO2-fixing enzymes) to redirect carbon from industrial effluents into useful bioproducts.

Oligothiophene compounds inhibit the membrane fusion between H5N1 avian influenza virus and the endosome of host cell.

Hemagglutinin (HA) which is essential for influenza viral infection and replication has become a target for the design of anti-influenza drugs. A novel series of oligothiophene compounds focused on the target were synthesized as specific inhibitors against the H5 subtype of influenza A viruses because oligothiophene has stronger π-π interactions with residues F1102 and M241 of HA2 side chains. Oligothiophene compounds were designed and synthesized by a series of alkylation, azidation, amination and amidation reactions. The entry inhibitory activities of those compounds were tested at a cellular level against H5N1 influenza pseudovirus. Compound 3sf was revealed as the most active inhibitor in this series with an IC50 of 0.029 µM. The activity of 3sf is almost 1000 times that of the positive reference compound (CL-385319). A structure-activity analysis of these compounds demonstrated that the size of the oligothiophene compounds was very important for the inhibitory activity. Four compounds (3sk, 3sf, 3sc and 4sc) of strong inhibitiory activity against H5N1 influenza pseudovirus were assessed against H1N1 influenza virus MDCK. They also showed strong inhibitiory activity with IC50s of 3.292 µM, 1.240 µM, 1.119 µM and 0.768 µM, respectively.

The self-assembly of a camptothecin-lysine nanotube.

A simple, low molecular weight camptothecin-lysine conjugate is reported to self-assemble into nanotubes with diameters of 70-100nm and a drug loading level of 60.5%. The nanotubes exhibited promising in vitro cytotoxicity against cancer cell lines A549, NCI-H460 and NCI-H23. The release of active camptothecin was highly dependent on conjugate concentration, temperature and pH of the solution.

Self-assembly of a 5-fluorouracil-dipeptide hydrogel.

The self-assembly of 5-fluorouracil dilysine conjugates into self-supporting hydrogels, comprised of entangled nanofibers or rigid nanotubes with diameters of 10 and 16 nm, respectively, is reported. The rate of release of 5-Fu from the conjugates was highly dependent on concentration in solution, whereas, release from the fully formed hydrogels was significantly slower. The 5-Fu conjugate also exhibited promising in vitro cytotoxicity against human tumor cell lines A549, H460 and H23.

A self-assembled nanotube for the direct aldol reaction in water.

Nanotubes formed by the aqueous assembly of a proline-lysine dipeptide (1) were used to create the hydrophobic microenvironments required to catalyze the aldol reaction in water. The self-assembly process occurred most efficiently in the presence of the substrates, producing an array of homogeneous nanotubes under the reaction conditions. The nanotubes formed by dipeptide 1 served as an efficient catalyst for the aldol reaction that functioned at low loading levels and provided good to excellent conversions. The catalytic activity of 1 was minimal under conditions that dissociated the nanotube into soluble monomers.