A Sustainable Biomineralization Approach for the Synthesis of Highly Fluorescent Ultra-Small Pt Nanoclusters.

Herein we report the first example of a facile biomineralization process to produce ultra-small-sized highly fluorescent aqueous dispersions of platinum noble metal quantum clusters (Pt-NMQCs) using a multi-stimulus responsive, biomimetic intrinsically disordered protein (IDP), Rec1-resilin. We demonstrate that Rec1-resilin acts concurrently as the host, reducing agent, and stabilizer of the blue-green fluorescent Pt-NMQCs once they are being formed. The photophysical properties, quantum yield, and fluorescence lifetime measurements of the synthesized Pt-NMQCs were examined using UV-Vis and fluorescence spectroscopy.

Agrocin 108 is a 5'-cytidine nucleotide bacteriocin containing a carbocyclic phosphoryl-ascorbate group.

Agrocin 108 is a 3'-O-β-D-xylopyranosyl-cytidine-5'-O-phosphodiester of an ascorbate-carbocyclic cyclopentenone analogue, with bacteriocin-like properties. This bacteriocin exhibits orders of magnitude greater than the inhibition zone diameter towards the indicator strain than either ampicillin or streptomycin. It has been isolated from cultures of Rhizobium rhizogenes strain K108.

Effects of Crowding and Environment on the Evolution of Conformational Ensembles of the Multi-Stimuli-Responsive Intrinsically Disordered Protein, Rec1-Resilin: A Small-Angle Scattering Investigation.

In this study, we explore the overall structural ensembles and transitions of a biomimetic, multi-stimuli-responsive, intrinsically disordered protein (IDP), Rec1-resilin. The structural transition of Rec1-resilin with change in molecular crowding and environment is evaluated using small-angle neutron scattering and small-angle X-ray scattering. The quantitative analyses of the experimental scattering data using a combination of computational models allowed comprehensive description of the structural evolution, organization, and conformational ensembles of Rec1-resilin in response to the changes in concentration, pH, and temperature.

A multi-responsive intrinsically disordered protein (IDP)-directed green synthesis of fluorescent gold nanoclusters.

Herein we demonstrate the green synthesis of fluorescent gold nanoclusters (AuNCs) using a multi-responsive intrinsically disordered protein (IDP) polymer, Rec1-resilin, as a multi-functional template. In a controlled environment, Rec1-resilin acts simultaneously as the directing agent and the reducer, and performs the role of a highly efficient stabilizer once AuNCs are formed. The evolution of the photophysical properties and the chemical states of AuNCs formed are measured using UV-Vis, fluorescence and X-ray photoelectron spectroscopy.

Structural ensembles reveal intrinsic disorder for the multi-stimuli responsive bio-mimetic protein Rec1-resilin.

Rec1-resilin is the first recombinant resilin-mimetic protein polymer, synthesized from exon-1 of the Drosophila melanogaster gene CG15920 that has demonstrated unusual multi-stimuli responsiveness in aqueous solution. Crosslinked hydrogels of Rec1-resilin have also displayed remarkable mechanical properties including near-perfect rubber-like elasticity. The structural basis of these extraordinary properties is not clearly understood.

Multi-responsive biomaterials and nanobioconjugates from resilin-like protein polymers.

Nature, through evolution over millions of years, has perfected materials with amazing characteristics and awe-inspiring functionalities that exceed the performance of man-made synthetic materials. One such remarkable material is native resilin - an extracellular skeletal protein that plays a major role in the jumping, flying, and sound production mechanisms in many insects. It is one of the most resilient (energy efficient) elastomeric biomaterials known with a resilience of ∼97% and a fatigue life in excess of 300 million cycles.

An16-resilin: an advanced multi-stimuli-responsive resilin-mimetic protein polymer.

Engineered protein polymers that display responsiveness to multiple stimuli are emerging as a promising class of soft material with unprecedented functionality. The remarkable advancement in genetic engineering and biosynthesis has created the opportunity for precise control over the amino acid sequence, size, structure and resulting functions of such biomimetic proteins. Herein, we describe the multi-stimuli-responsive characteristics of a resilin-mimetic protein, An16-resilin (An16), derived from the consensus sequence of resilin gene in the mosquito Anopheles gambiae.

Identification of proteins associated with adhesive prints from Holothuria dofleinii Cuvierian tubules.

Cuvierian tubules are expelled as a defence mechanism against predators by various species within the family Holothuridae. When the tubules are expelled, they become sticky almost immediately and ensnare the predator. The mechanism of this rapid adhesion is not clear, but proteins on the surface of the expelled tubules are widely believed to be involved.

Controlled surface modification of tissue culture polystyrene for selective cell binding using resilin-inspired polypeptides.

Modified tissue culture polystyrene (TCP) surfaces have been fabricated by attachment of recombinant polypeptides based on Drosophila melanogaster resilin and the Anopheles gambiae resilin-like protein. The D. melanogaster polypeptide (Rec-1) was from the first exon of resilin and consisted of 17 very similar repeats of a 15 residue sequence.

Purification of recombinant protein by cold-coacervation of fusion constructs incorporating resilin-inspired polypeptides.

Polypeptides containing between 4 and 32 repeats of a resilin-inspired sequence AQTPSSYGAP, derived from the mosquito Anopheles gambiae, have been used as tags on recombinant fusion proteins. These repeating polypeptides were inspired by the repeating structures that are found in resilins and sequence-related proteins from various insects. Unexpectedly, an aqueous solution of a recombinant resilin protein displays an upper critical solution temperature (cold-coacervation) when held on ice, leading to a separation into a protein rich phase, typically exceeding 200 mg/mL, and a protein-poor phase.

Stabilization of collagen tissues by photocrosslinking.

Photocrosslinking, using 2 mM Ru(II)(bpy)(3)Cl(2) and various concentrations of sodium persulfate with irradiation by blue light, ∼455 nm, has been shown to be a rapid and effective method for crosslinking various tissues: tendon, amnion membrane, pericardium, and heart valve leaflet. The presence of new crosslinking was demonstrated by the increase in the shrinkage temperature of these tissues. In all the cases, increase in the shrinkage temperatures were seen, although at higher sodium persulfate concentrations, for example, 100 mM, both with and without the Ru(II)(bpy)(3)Cl(2) catalyst, some degradation of the collagenous tissues was found.

Expression of the rubber-like protein, resilin, in developing and functional insect cuticle determined using a Drosophila anti-Rec 1 resilin antibody.

Background: The natural elastomeric protein, insect resilin, is the most efficient elastic material known, used to store energy for jumping and flight in a variety of insects. Here, an antibody to recombinant Drosophila melanogaster pro-resilin is used to examine resilin expression in Drosophila and a wider range of insects.

Results: Immunostaining of Drosophila embryos reveals anti-resilin reactivity in epidermal patches that exhibit a dynamic spatial and temporal expression through late embryogenesis.

Antibody labelling of resilin in energy stores for jumping in plant sucking insects.

The rubbery protein resilin appears to form an integral part of the energy storage structures that enable many insects to jump by using a catapult mechanism. In plant sucking bugs that jump (Hemiptera, Auchenorrhyncha), the energy generated by the slow contractions of huge thoracic jumping muscles is stored by bending composite bow-shaped parts of the internal thoracic skeleton. Sudden recoil of these bows powers the rapid and simultaneous movements of both hind legs that in turn propel a jump.

A highly elastic and adhesive gelatin tissue sealant for gastrointestinal surgery and colon anastomosis.

Background: We describe the development of a highly elastic and adhesive surgical tissue sealant, based on photochemically crosslinked gelatin, for sealing sutured incisions in the gastrointestinal (GI) tract in a rabbit surgical model and in a canine colon anastomosis study.

Methods: The study included in vitro assessment of mechanical parameters of the tissue sealant and in vivo analysis of burst strength and histology at 24 h, 3 days and 7 days post surgery, in a rabbit model, to assess progress of wound healing at the suture sites. Utility of this sealant to repair and seal a lower colonic resection and anastomosis procedure in a canine model was also investigated.

Molecular and functional characterisation of resilin across three insect orders.

Resilin is an important elastomeric protein of insects, with roles in the storage and release of energy during a variety of different functional categories including flight and jumping. To date, resilin genes and protein function have been characterised only in a small number of flying insects, despite their importance in fleas and other jumping insects. Microscopy and immunostaining studies of resilin in flea demonstrate the presence of resilin pads in the pleural arch at the top of the hind legs, a region responsible for the flea's jumping ability.

The effect of hydration on molecular chain mobility and the viscoelastic behavior of resilin-mimetic protein-based hydrogels.

The outstanding rubber-like elasticity of resilin and resilin-mimetic proteins depends critically on the level of hydration. In this investigation, water vapor sorption and the role of hydration on the molecular chain dynamics and viscoelastic properties of resilin-mimetic protein, rec1-resilin is investigated in detail. The dynamic and equilibrium swelling behavior of the crosslinked protein hydrogels with different crosslink density are reported under various controlled environments.

Self-organization, interfacial interaction and photophysical properties of gold nanoparticle complexes derived from resilin-mimetic fluorescent protein rec1-resilin.

In this investigation we report the synthesis of optically coupled hybrid architectures based on a new biomimetic fluorescent protein rec1-resilin and nanometer-scale gold nanoparticles (AuNPs) in a one-step method using a non-covalent mode of binding protocol. The presence of uniformly distributed fluorophore sequences, -Ser(Thr)-Tyr-Gly- along the molecular structure of rec1-resilin provides significant opportunity to synthesize fluorophore-modified AuNPs bioconjugates with unique photophysical properties. The detailed analyses of the AuNP-bioconjugates, synthesized under different experimental conditions using spectroscopic, microscopic and scattering techniques demonstrate the organizational pathways and the electronic and photophysical properties of the developed AuNP-rec1-resilin bioconjugates.

Photochemical crosslinking of soluble wool keratins produces a mechanically stable biomaterial that supports cell adhesion and proliferation.

Keratins extracted from various "hard tissues" such as wool, hair, and nails are increasingly being investigated as a source of abundant, biocompatible materials. In this study we explored a recent photochemical method to crosslink solubilized wool keratoses, with the aim of producing a mechanically favorable biomaterial. Wool proteins were isolated by oxidizing the disulfides and extracting the resulting soluble keratoses.

Photochemically crosslinked matrices of gelatin and fibrinogen promote rapid cell proliferation.

Here we report the use of a facile photochemical crosslinking method to fabricate stable polymer matrices from unmodified gelatin and fibrinogen. Gels were produced by covalent crosslinking of the proteins in a rapid photo-oxidative process, catalysed by a ruthenium metal complex and irradiation with visible light. For generation of macroporous, spongy matrices, the proteins and crosslinking reagents were mixed with catalase and hydrogen peroxide to achieve a foaming reaction, producing a stable, foamed matrix that was subsequently photo-crosslinked.

A highly elastic tissue sealant based on photopolymerised gelatin.

Gelatin is widely used as a medical biomaterial because it is readily available, cheap, biodegradable and demonstrates favourable biocompatibility. Many applications require stabilisation of the biomaterial by chemical crosslinking, and this often involves derivatisation of the protein or treatment with cytotoxic crosslinking agents. We have previously shown that a facile photochemical method, using blue light, a ruthenium catalyst and a persulphate oxidant, produces covalent di-tyrosine crosslinks in resilin and fibrinogen to form stable hydrogel biomaterials.

A pH-responsive interface derived from resilin-mimetic protein Rec1-resilin.

In this investigation, for the first time we report the effects of pH on the molecular orientation, packing density, structural properties, adsorption characteristics and viscoelastic behaviour of resilin-mimetic protein rec1-resilin at the solid-liquid interface using quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) spectroscopy. QCM-D and SPR data confirm that the binding ability of rec1-resilin on a substrate is strongly pH-dependent the protein packing density on a gold surface is calculated to be 4.45 x 10(13) per cm(2) at the isoelectric point (IEP approximately 4.

Comparisons of recombinant resilin-like proteins: repetitive domains are sufficient to confer resilin-like properties.

Two novel recombinant proteins An16 and Dros16 have recently been generated. These recombinant proteins contain, respectively, sixteen copies of an 11 amino acid repetitive domain (AQTPSSQYGAP) observed in a resilin-like gene from Anopheles gambiae and sixteen copies of a 15 amino acid repetitive domain (GGRPSDSYGAPGGGN) observed in the first exon of the Drosophila melanogaster CG15920 gene. We compare structural characteristics of the proteins and material properties of resulting biopolymers relative to Rec1-resilin, a previously characterized resilin-like protein encoded by the first exon of the Drosophila melanogaster CG15920 gene.

Physical approaches for fabrication of organized nanostructure of resilin-mimetic elastic protein rec1-resilin.

Protein adsorption on surfaces is a fundamental step in many applications. While various methods such as lithography, self assembly using nanoparticles, layer-by-layer attachment, etc. have been employed, here we report fabrication of controlled nanostructure of a new resilin-mimetic elastic protein rec1-resilin using physical approaches.

The development of photochemically crosslinked native fibrinogen as a rapidly formed and mechanically strong surgical tissue sealant.

We recently reported the generation of a highly elastic, crosslinked protein biomaterial via a rapid photochemical process using visible light illumination. In light of these findings, we predicted that other unmodified, tyrosine-rich, self-associating proteins might also be susceptible to this covalent crosslinking method. Here we show that unmodified native fibrinogen can also be photochemically crosslinked into an elastic hydrogel biomaterial through the rapid formation of intermolecular dityrosine.