Protein-based materials, toward a new level of structural control

Through billions of years of evolution nature has created and refined structural proteins for a wide variety of specific purposes. Amino acid sequences and their associated folding patterns combine to create elastic, rigid or tough materials. In many respects, nature’s intricately designed products provide challenging examples for materials scientists, but translation of natural structural concepts into bio-inspired materials requires a level of control of macromolecular architecture far higher than that afforded by conventional polymerization processes. An increasingly important approach to this problem has been to use biological systems for production of materials. Through protein engineering, artificial genes can be developed that encode protein-based materials with desired features. Structural elements found in nature, such as β-sheets and α-helices, can be combined with great flexibility, and can be outfitted with functional elements such as cell binding sites or enzymatic domains. The possibility of incorporating non-natural amino acids increases the versatility of protein engineering still further. It is expected that such methods will have large impact in the field of materials science, and especially in biomedical materials science, in the future

Biomimetic spatial and temporal (4D) design and fabrication

We imagine the built environment of the future as a ‘bio-hybrid machine for living in’ that will sense and react to activities within the space in order to provide experiences and services that will elevate quality of life while coexisting seamlessly with humans and the natural environment. The study of Hierarchical design in biological materials has the potential to alter the way designers/ engineers/ crafts-men of the future engage with materials in order to realise such visions. We are ex-ploring this design approach using digital manufacturing technologies such as jac-quard weaving and 3D printing

Dynamics of Phase Behavior of a Polymer Blend Under Shear Flow

We study the dynamics of the phase behavior of a polymer blend in the presence of shear flow. By adopting a two fluid picture and using a generalization of the concept of material derivative, we construct kinetic equations that describe the phase behavior of polymer blends in the presence of external flow. A phenomenological form for the shear modulus for the blend is proposed. The study indicates that a nonlinear dependence of the shear modulus of the blend on the volume fraction of one of the species is crucial for a shift in the stability line to be induced by shear flow.Comment: 16 pages, late

EPR Study of Spin Labeled Brush Polymers in Organic Solvents

Spin-labeled polylactide brush polymers were synthesized via ring-opening metathesis polymerization (ROMP), and nitroxide radicals were incorporated at three different locations of brush polymers: the end and the middle of the backbone, and the end of the side chains (periphery). Electron paramagnetic resonance (EPR) was used to quantitatively probe the macromolecular structure of brush polymers in dilute solutions. The peripheral spin-labels showed significantly higher mobility than the backbone labels, and in dimethylsulfoxide (DMSO), the backbone end labels were shown to be more mobile than the middle labels. Reduction of the nitroxide labels by a polymeric reductant revealed location-dependent reactivity of the nitroxide labels: peripheral nitroxides were much more reactive than the backbone nitroxides. In contrast, almost no difference was observed when a small molecule reductant was used. These results reveal that the dense side chains of brush polymers significantly reduce the interaction of the backbone region with external macromolecules, but allow free diffusion of small molecules

State-Selective Metabolic Labeling of Cellular Proteins

Transcriptional activity from a specified promoter can provide a useful marker for the physiological state of a cell. Here we introduce a method for selective tagging of proteins made in cells in which specified promoters are active. Tagged proteins can be modified with affinity reagents for enrichment or with fluorescent dyes for visualization. The method allows state-selective analysis of the proteome, whereby proteins synthesized in predetermined physiological states can be identified. The approach is demonstrated by proteome-wide labeling of bacterial proteins upon activation of the P_(BAD) promoter and the SoxRS regulon and provides a basis for analysis of more complex systems including spatially heterogeneous microbial cultures and biofilms

Core-Clickable PEG-Branch-Azide Bivalent-Bottle-Brush Polymers by ROMP: Grafting-Through and Clicking-To

The combination of highly efficient polymerizations with modular "click" coupling reactions has enabled the synthesis of a wide variety of novel nanoscopic tructures. Here we demonstrate the facile synthesis of a new class of clickable, branched nanostructures, polyethylene glycol (PEG)-branch-azide bivalent-brush polymers, facilitated by "graft-through" ring-opening metathesis polymerization of a branched norbornene-PEG-chloride macromonomer followed by halide-azide exchange. The resulting bivalent-brush polymers possess azide groups at the core near a polynorbornene backbone with PEG chains extended into solution; the structure resembles a unimolecular micelle. We demonstrate copper-catalyzed azide-alkre cycloaddition (CuAAC) "click-to" coupling of a photocleavable doxorubicin (DOX)-alkyne derivative to the azide core. The CuAAC coupling was quantitative across a wide range of nanoscopic sizes (similar to 6-similar to 50 nrn); UV photolysis of the resulting DOX-loaded materials yielded free DOX that was therapeutically effective against human cancer cells

Thermal and Structural Properties of Biologically Derived Monodisperse Hairy-Rod Polymers

Monodisperse derivatives of poly(γ-4-(hexadecyloxy)benzyl α,l-glutamate) (PHBG-X, X = 3 or 4) with backbone sequence GluAsp(Glu_(17)Asp)_xGluGlu were prepared by reaction of 4-(hexadecyloxy)phenyldiazomethane with the corresponding monodisperse poly(α,l-glutamate) (PLGA) derivatives (PLGA-X, X = 3 or 4). PHBG-3 and -4 exhibited strong endotherms near 45 °C and weak endotherms near 86 °C when analyzed by differential scanning calorimetry. X-ray diffraction suggested that these polymers aggregate to form layerlike solid structures at room temperature, with extended alkyl side chains forming paraffinlike crystallites. Most of the side chain order disappears at the first melting transition; however, the layerlike structure remains. Both polymers are isotropic above the second melting transition; no ordered melts were observed at higher temperatures, possibly due to the small aspect ratios of PHBG-3 and -4. In contrast, polydisperse poly(γ-4-(hexadecyloxy)benzyl α,l-glutamate) (PDI = 1.2, DP = 98) (PHBG-P1), prepared from commercial PLGA, formed liquid crystalline (LC) phases between 97 and 105 °C

Internal segregation and side chain ordering in hairy-rod polypeptide monolayers at the gas/water interface: An x-ray scattering study

We report studies of the structure and packing of Langmuirmonolayers (LMs) of polypeptide poly(γ-4-(n-hexadecyloxy)benzyl α,L-glutamate) (C16–O–PBLG) on the surface of water. The molecule is a “hairy rod” and consists of side attachments of hexadecyloxy chains (–O–C16) to the rigid rod-like core made up of α-helical poly(γ-benzyl L-glutamate) (PBLG). Measurements include surface pressure (Π) versus area/monomer (A) isotherms, x-ray specular reflectivity (XR), and grazing incidence diffraction(GID). In contrast to the LM of bare PBLG on water, which undergoes a monolayer/bilayer transition with increasing Π, monolayers of C16–O–PBLG remain stable up to the highest densities. On the basis of XR and GID results, the structure of the C16–O–PBLG monolayer is characterized by the following main features. First, hydrophobicity causes the –O–C16 chains to segregate towards the film/gas interface and away from water and the PBLG cores, which sit parallel to and near the water/film interface. Since the attachment position of some of the side chains is at the core/water interface, the segregation forces these chains into the space between neighboring core rods. Compression associated with increasing Π thickens the film but the internally segregated structure is maintained for all Π (i.e., >∼30 dyne/cm). Second, the C16–O–PBLG rods form domains in which the rods are aligned parallel to each other and to the interface. The correlation length for the interhelix positional order of the rods is short and typically comparable to or less than the length of the rods. With increasing Π the spacing d between nearest-neighbor rods decreases linearly with A at high Π, indicating a direct correspondence between the macroscopic compressibility and the microscopic interhelix compressibility. Third, as Π increases past ∼5 dyne/cm, the local packing of tethered –O–C16 chains displays the same herringbone (HB) order that is common for high-density bulk and monolayer phases of alkyl chains. Various features of the observed GID peaks also imply that the HB order of –O–C16 chains is oriented with respect to the helical axes of aligned PBLG cores. We propose that the HB order is established initially by one-dimensionally confined chains between aligned rods at low Π and grows laterally with compression

Internal Segregation and Side Chain Ordering in Hairy-Rod Polypeptide Monolayers at the Gas/Water Interface: An X-Ray Scattering Study

We report studies of the structure and packing of Langmuir monolayers (LMs) of polypeptide poly($\gamma$-4-(n-hexadecyloxy)benzyl $\alpha$,L-glutamate) (C16–O–PBLG) on the surface of water. The molecule is a “hairy rod” and consists of side attachments of hexadecyloxy chains (–O–C16) to the rigid rod-like core made up of $\alpha$-helical poly($\gamma$-benzyl L-glutamate) (PBLG). Measurements include surface pressure ($\Pi$) versus area/monomer (A) isotherms, x-ray specular reflectivity (XR), and grazing incidence diffraction (GID). In contrast to the LM of bare PBLG on water, which undergoes a monolayer/bilayer transition with increasing $\Pi$, monolayers of C16–O–PBLG remain stable up to the highest densities. On the basis of XR and GID results, the structure of the C16–O–PBLG monolayer is characterized by the following main features. First, hydrophobicity causes the –O–C16 chains to segregate towards the film/gas interface and away from water and the PBLG cores, which sit parallel to and near the water/film interface. Since the attachment position of some of the side chains is at the core/water interface, the segregation forces these chains into the space between neighboring core rods. Compression associated with increasing $\Pi$ thickens the film but the internally segregated structure is maintained for all $\Pi$ (i.e., >$\sim$30 dyne/cm). Second, the C16–O–PBLG rods form domains in which the rods are aligned parallel to each other and to the interface. The correlation length for the interhelix positional order of the rods is short and typically comparable to or less than the length of the rods. With increasing $\Pi$ the spacing d between nearest-neighbor rods decreases linearly with A at high $\Pi$, indicating a direct correspondence between the macroscopic compressibility and the microscopic interhelix compressibility. Third, as $\Pi$ increases past $\sim$5 dyne/cm, the local packing of tethered –O–C16 chains displays the same herringbone (HB) order that is common for high-density bulk and monolayer phases of alkyl chains. Various features of the observed GID peaks also imply that the HB order of –O–C16 chains is oriented with respect to the helical axes of aligned PBLG cores. We propose that the HB order is established initially by one-dimensionally confined chains between aligned rods at low Π and grows laterally with compression.Engineering and Applied Science

Structure and Dynamics of Hybrid Colloid-Polyelectrolyte Coacervates: Insights from Molecular Simulations

Electrostatic interactions in polymeric systems are responsible for a wide range of liquid-liquid phase transitions that are of importance for biology and materials science. Such transitions are referred to as complex coacervation, and recent studies have sought to understand the underlying physics and chemistry. Most theoretical and simulation efforts to date have focused on oppositely charged linear polyelectrolytes, which adopt nearly ideal-coil conformations in the condensed phase. However, when one of the coacervate components is a globular protein, a better model of complexation should replace one of the species with a spherical charged particle or colloid. In this work, we perform coarse-grained simulations of colloid-polyelectrolyte coacervation using a spherical model for the colloid. Simulation results indicate that the electroneutral cell of the resulting (hybrid) coacervates consists of a polyelectrolyte layer adsorbed on the colloid. Power laws for the structure and the density of the condensed phase, which are extracted from simulations, are found to be consistent with the adsorption-based scaling theory of coacervation. The coacervates remain amorphous (disordered) at a moderate colloid charge, $Q$, while an intra-coacervate colloidal crystal is formed above a certain threshold, at $Q > Q^{*}$. In the disordered coacervate, if $Q$ is sufficiently low, colloids diffuse as neutral non-sticky nanoparticles in the semidilute polymer solution. For higher $Q$, adsorption is strong and colloids become effectively sticky. Our findings are relevant for the coacervation of polyelectrolytes with proteins, spherical micelles of ionic surfactants, and solid organic or inorganic nanoparticles