Temporally Programmed Disassembly and Reassembly of C3Ms.

Responsive materials, which can adapt and operate autonomously under dynamic conditions, are a stepping stone towards functional, life-like systems inspired by fueled self-assembly processes in nature. Complex coacervate core micelles (C3Ms) comprising oppositely charged macromolecules constitute a novel class of polymeric micelles ideally suited for use as responsive nanoscopic delivery vehicles of hydrophilic and hydrophobic cargo. To fully exploit their potential, it is important that the C3Ms form and fall apart in an autonomous fashion as orchestrated by dynamic cues in their environment. Herein a means to temporally program the self-regulated C3M coassembly pathway, using a modulated base-catalyzed feedback system, is presented. Incorporated in the C3Ms is a pH responsive polyfluorene-based conjugated polyelectrolyte (CPF) as a building block and trace amounts of a molecular sensor (doxorubicin HCl) as cargo, both of which report on micellar coassembly and disassembly via binding-induced fluorescence quenching. CPF additionally reports on the pH of its microenvironment as its pH-dependent conformational states are mirrored in the transitions of its vibronic bands. This experimental design enables one to monitor solution pH, C3M disassembly and reassembly, as well as cargo release and recapture noninvasively in a closed system with real time florescence experiments.

Complex Coacervate Core Micelles Containing Poly(vinyl alcohol) Inhibit Ice Recrystallization.

Complex coacervate core micelles (C3Ms) form upon complexation of oppositely charged copolymers. These co-assembled structures are widely investigated as promising building blocks for encapsulation, nanoparticle synthesis, multimodal imaging, and coating technology. Here, the impact on ice growth is investigated of C3Ms containing poly(vinyl alcohol), PVA, which is well known for its high ice recrystallization inhibition (IRI) activity. The PVA-based C3Ms are prepared upon co-assembly of poly(4-vinyl-N-methyl-pyridinium iodide) and poly(vinyl alcohol)-block-poly(acrylic acid). Their formation conditions, size, and performance as ice recrystallization inhibitors are studied. It is found that the C3Ms exhibit IRI activity at PVA monomer concentrations as low as 1 × 10-3 m. The IRI efficacy of PVA-C3Ms is similar to that of linear PVA and PVA graft polymers, underlining the influence of vinyl alcohol monomer concentration rather than polymer architecture.

Illuminating the Reaction Pathways of Viromimetic Assembly.

The coassembly of well-defined biological nanostructures relies on a delicate balance between attractive and repulsive interactions between biomolecular building blocks. Viral capsids are a prototypical example, where coat proteins exhibit not only self-interactions but also interact with the cargo they encapsulate. In nature, the balance between antagonistic and synergistic interactions has evolved to avoid kinetic trapping and polymorphism. To date, it has remained a major challenge to experimentally disentangle the complex kinetic reaction pathways that underlie successful coassembly of biomolecular building blocks in a noninvasive approach with high temporal resolution. Here we show how macromolecular force sensors, acting as a genome proxy, allow us to probe the pathways through which a viromimetic protein forms capsids. We uncover the complex multistage process of capsid assembly, which involves recruitment and complexation, followed by allosteric growth of the proteinaceous coat. Under certain conditions, the single-genome particles condense into capsids containing multiple copies of the template. Finally, we derive a theoretical model that quantitatively describes the kinetics of recruitment and growth. These results shed new light on the origins of the pathway complexity in biomolecular coassembly.

Stark fluorescence spectroscopy reveals two emitting sites in the dissipative state of FCP antennas.

Diatoms are characterized by very efficient photoprotective mechanisms where the excess energy is dissipated as heat in the main antenna system constituted by fucoxanthin-chlorophyll (Chl) protein complexes (FCPs). We performed Stark fluorescence spectroscopy on FCPs in their light-harvesting and energy dissipating states. Our results show that two distinct emitting bands are created upon induction of energy dissipation in FCPa and possibly in FCPb. More specifically one band is characterized by broad red shifted emission above 700nm and bears strong similarity with a red shifted band that we detected in the dissipative state of the major light-harvesting complex II (LHCII) of plants [26]. We discuss the results in the light of different mechanisms proposed to be responsible for photosynthetic photoprotection.

Monitoring Protein Capsid Assembly with a Conjugated Polymer Strain Sensor.

Semiconducting polymers owe their optoelectronic properties to the delocalized electronic structure along their conjugated backbone. Their spectral features are therefore uniquely sensitive to the conformation of the polymer, where mechanical stretching of the chain leads to distinct vibronic shifts. Here we demonstrate how the optomechanical response of conjugated polyelectrolytes can be used to detect their encapsulation in a protein capsid. Coating of the sensor polymers by recombinant coat proteins induces their stretching due to steric hindrance between the proteins. The resulting mechanical planarizations lead to pronounced shifts in the vibronic spectra, from which the process of capsid formation can be directly quantified. These results show how the coupling between vibronic states and mechanical stresses inherent to conjugated polymers can be used to noninvasively measure strains at the nanoscale.

Equivalent pathways in melting and gelation of well-defined biopolymer networks.

We use multiple particle tracking microrheology to study the melting and gelation behavior of well-defined collagen-inspired designer biopolymers expressed by the transgenic yeast P. Pastoris. The system consists of a hydrophilic random coil-like middle block and collagen-like end block. Upon cooling, the end blocks assemble into well-defined transient nodes with exclusively 3-fold functionality. We apply the method of time-cure superposition of the mean-square displacement of tracer beads embedded in the biopolymer matrix to study the kinetics and thermodynamics of approaching the gel point from both the liquid and the solid side. The melting point, gel point, and critical relaxation exponents are determined from the shift factors of the mean-square displacement and we discuss the use of dynamic scaling exponents to correctly determine the critical transition. Critical relaxation exponents obtained for different concentrations in both systems are compared with the currently existing dynamic models in literature. In our study, we find that, while the time scales of gelation and melting are different by orders of magnitude, and show inverse dependence on concentration, that the pathways followed are completely equivalent.

Effect of UV Irradiation Time and Headgroup Interactions on the Reversible Colorimetric pH Response of Polydiacetylene Assemblies.

Polydiacetylenes are chromatic conjugated polymers formed upon the photopolymerization of self-assembled diacetylenes. They exhibit conformation-dependent colorimetric responses, usually irreversible, to external triggers. Here, we presented an approach to obtain a reversible colorimetric response to a pH change through structural modifications on the monomer and extended photopolymerization time. Both factors, enhanced hydrogen bond forming headgroups and longer UV exposure, impacted the rotational freedom of polydiacetylene conformation. Such a restricted conformation state reduced colorimetric response efficiency but enabled reversible colorimetric response to a pH change. These results highlight the possibility of obtaining a reversible colorimetric pH response of polydiacetylenes for customized sensing applications through monomer-level tailoring combined with tuning the photopolymerization time.