Resilin is a member of a family of elastic proteins that includes elastin, as well as gluten, gliadin, abductin and spider silks. Resilin is found in specialized regions of the cuticle of most insects, providing low stiffness, high strain and efficient energy storage; it is best known for its roles in insect flight and the remarkable jumping ability of fleas and spittle bugs. Previously, the Drosophila melanogaster CG15920 gene was tentatively identified as one encoding a resilin-like protein (pro-resilin). Here we report the cloning and expression of the first exon of the Drosophila CG15920 gene as a soluble protein in Escherichia coli. We show that this recombinant protein can be cast into a rubber-like biomaterial by rapid photochemical crosslinking. This observation validates the role of the putative elastic repeat motif in resilin function. The resilience (recovery after deformation) of crosslinked recombinant resilin was found to exceed that of unfilled synthetic polybutadiene, a high resilience rubber. We believe that our work will greatly facilitate structural investigations into the functional properties of resilin and shed light on more general aspects of the structure of elastomeric proteins. In addition, the ability to rapidly cast samples of this biomaterial may enable its use in situ for both industrial and biomedical applications.
Summary 1.Ungulates have reached such high densities in some natural ecosystems that culling is frequently used to reduce their impacts on vegetation. However, much is still unknown about the outcomes of landscape-level control, in part because monitoring vegetation recovery requires decades. 2. We report long-term vegetation changes in permanent plots located in forest, shrubland and grassland communities across a mountain range in southern New Zealand. We test whether c. 92% reduction in the population of invasive non-indigenous red deer Cervus elaphus since 1964 has led to the recovery of deer-preferred species. 3. Tree seedlings, saplings and the number of seedlings per adult tree increased over time. There was lower recruitment, however, of highly palatable forest species compared with less palatable species, and the recruitment of saplings was lower in browsed forest plots compared with deer exclosures. 4. The total number of occurrences and absolute number of palatable species per plot increased over time in shrublands and grasslands respectively. The height of both shrublands and palatable grassland snow tussocks Chionochloa spp. increased, although the occurrences of most individual species remained unchanged over time. 5. Vegetation recovery at our site in response to long-term and significant herbivore reductions may be limited by several factors, including the slow growth rates of New Zealand species, densitydependent diet switching by deer, altered successional trajectories and below-ground processes. 6. Synthesis and applications. Our results suggest that after nearly four decades, even low densities of introduced herbivores may restrict ecosystem recovery, and therefore, restoring herbivore-disturbed ecosystems by solely manipulating herbivore population numbers may require a long-term perspective. Management strategies can accelerate recovery by protecting existing palatable plants within deer-exclosures, and planting or seeding palatable species within these refugia. However, in addition to increasing seed sources, restoration may only become apparent following large-scale disturbance events and canopy turnover.
When provoked, Notaden bennetti frogs secrete an exudate which rapidly forms a tacky elastic solid ("frog glue"). This protein-based material acts as a promiscuous pressure-sensitive adhesive that functions even in wet conditions. We conducted macroscopic tests in air to assess the tensile strength of moist glue (up to 78 +/- 8 kPa) and the shear strength of dry glue (1.7 +/- 0.3 MPa). We also performed nanomechanical measurements in water to determine the adhesion (1.9-7.2 nN or greater), resilience (43-56%), and elastic modulus (170-1035 kPa) of solid glue collected in different ways. Dry glue contains little carbohydrate and consists mainly of protein. The protein complement is rich in Gly (15.8 mol %), Pro (8.8 mol %), and Glu/Gln (14.1 mol %); it also contains some 4-hydroxyproline (4.6 mol %) but no 5-hydroxylysine or 3,4-dihydroxyphenylalanine (L-Dopa). Denaturing gel electrophoresis of the glue reveals a characteristic pattern of proteins spanning 13-400 kDa. The largest protein (Nb-1R, apparent molecular mass 350-500 kDa) is also the most abundant, and this protein appears to be the key structural component. The solid glue can be dissolved in dilute acids; raising the ionic strength causes the glue components to self-assemble spontaneously into a solid which resembles the starting material. We describe scattering studies on dissolved and solid glue and provide microscopy images of glue surfaces and sections, revealing a porous interior that is consistent with the high water content (85-90 wt %) of moist glue. In addition to compositional similarities with other biological adhesives and well-known elastomeric proteins, the circular dichroism spectrum of dissolved glue is almost identical to that for soluble elastin and electron and scanning probe microscopy images invite comparison with silk fibroins. Covalent cross-linking does not seem to be necessary for the glue to set.
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