Segmented nanofibers of spider dragline silk: atomic force microscopy and single-molecule force spectroscopy. (65/701)

Despite its remarkable materials properties, the structure of spider dragline silk has remained unsolved. Results from two probe microscopy techniques provide new insights into the structure of spider dragline silk. A soluble synthetic protein from dragline silk spontaneously forms nanofibers, as observed by atomic force microscopy. These nanofibers have a segmented substructure. The segment length and amino acid sequence are consistent with a slab-like shape for individual silk protein molecules. The height and width of nanofiber segments suggest a stacking pattern of slab-like molecules in each nanofiber segment. This stacking pattern produces nano-crystals in an amorphous matrix, as observed previously by NMR and x-ray diffraction of spider dragline silk. The possible importance of nanofiber formation to native silk production is discussed. Force spectra for single molecules of the silk protein demonstrate that this protein unfolds through a number of rupture events, indicating a modular substructure within single silk protein molecules. A minimal unfolding module size is estimated to be around 14 nm, which corresponds to the extended length of a single repeated module, 38 amino acids long. The structure of this spider silk protein is distinctly different from the structures of other proteins that have been analyzed by single-molecule force spectroscopy, and the force spectra show correspondingly novel features.  (+info)

A new assay for the detection of Loxosceles species (brown recluse) spider venom. (66/701)

STUDY OBJECTIVE: Dermal lesions from unrelated arthropod species and medical causes appear similar to Loxosceles species (brown recluse spider) bites. This may result in delayed diagnosis and treatment. We developed a sensitive Loxosceles species venom enzyme-linked immunosorbent assay (ELISA) and characterized the specificity of the assay by evaluating antigenic cross-reactivity from a variety of North American arthropod venoms. METHODS: North American arthropod (14 spiders, 2 scorpions, and 1 bee) venoms were studied. Three venom amounts (diluted in 100 microL of ELISA buffer) were assayed: 16,000 ng, 2,000 ng, and 40 ng. The latter quantity was selected because this is the observed maximum amount of venom we detect when inoculating dermis with amounts likely to be deposited by a spider bite. The larger venom amounts are overwhelming quantities designed to test the limits of the assay for arthropod venom cross-reactivity. Similar amounts of Loxosceles species venom and bovine albumin served as positive and negative controls, respectively. RESULTS: At the lowest amount of venom tested (40 ng), the ELISA detected only the Loxosceles species positive control. When 2,000 ng was assayed, only Scytodes fusca and Kukulcania hibernalis arachnid venoms (in addition to Loxosceles species) cross-reacted to the assay. Finally, at 16,000 ng, the ELISA assay modestly detected Diguetia canities, Heteropoda venatoria, Tegenaria agrestis, Plectreurys tristes, Dolomedes tenebrosus, and Hadrurus arizonensis arachnid venoms. CONCLUSION: Cross-reactivity was observed in 8 of 17 North American arthropod venoms when large venom amounts were assayed with a Loxosceles species ELISA. By using a relevant quantity of venom, 40 ng, the assay was specific for Loxosceles species venom. The venom specificity of the ELISA may allow clinical application in Loxosceles species endemic regions of North America.  (+info)

Nuptial gifts of male spiders function as sensory traps. (67/701)

While nuptial food gifts come in various forms in arthropods, their evolutionary origins are unclear. A previous study on insects has shown that such gifts may arise as a sensory trap that exploits a female's underlying motivation to feed. Here I present independent evidence of a sensory trap in spiders. In certain visually oriented spiders, I suggest that males initially exploit the maternal care instinct by producing a nuptial gift that closely resembles the female egg sac. Males of the spider Pisaura mirabilis cover their prey gift with a silk layer, transforming it into a white round object. In a laboratory experiment I tested whether the colour of the gift affected the rate that females accepted males displaying their gifts. I found that the brighter and the more alike the nuptial gift to a female's egg sac, the faster the female responded by grabbing the gift. My results support the hypothesis that the nuptial gift in P. mirabilis works as a sensory trap.  (+info)

Influence of cues from the anterior medial eyes of virtual prey on Portia fimbriata, an araneophagic jumping spider. (68/701)

Portia fimbriata from Queensland, Australia, is a jumping spider (Salticidae) that preys on other spiders, including other salticids. Cryptic stalking (palps retracted, walking very slowly and freezing when faced) is a prey-specific tactic deployed exclusively against salticid prey. Using vision alone, P. fimbriata discriminates salticid from non-salticid prey, with the prey salticid's large anterior median (AM) eyes providing critical cues. Here, using computer-rendered virtual three-dimensional lures, we clarify experimentally some of the specific optical cues from AM eyes that influence the behaviour of P. fimbriata. Control lures were based on Jacksonoides queenslandicus, the salticid upon which P. fimbriata most commonly preys in nature. Experimental lures were modified to isolate specific combinations of AM eye features. For presentation to P. fimbriata, lures were projected on a small screen positioned in front of a web-covered platform. Each individual P. fimbriata was tested once with a control lure and once with a lure on which some AM eye feature had been altered. P. fimbriata was affected by the presence-versus-absence of AM eyes, by the size and shape of these eyes and by AM eye position. Horizontal position on the face of a single AM eye had no discernible influence on whether P. fimbriata initiated cryptic stalking, but reduced how often P. fimbriata froze when faced by the prey. The implications of the findings are discussed in relation to perceptual processes underlying the predatory strategy of P. fimbriata.  (+info)

Electrophysiological characterization and molecular identification of the Phoneutria nigriventer peptide toxin PnTx2-6. (69/701)

A cDNA with 403 nucleotides encoding the precursor of the toxin PnTx2-6 was cloned and sequenced. Subsequent analysis revealed that the precursor begins with a signal peptide and a glutamate-rich propeptide. The succeeding peptide confirmed the reported sequence of PnTx2-6. The purified toxin exerted complex effects on Na(+) current of frog skeletal muscle. There was a marked decrease of the inactivation kinetics, and a shift to hyperpolarizing potentials of both the Na(+) conductance and the steady-state inactivation voltage dependences, along with a reduction of the current amplitude. The concentration dependence of the modified current suggests a K(D) of 0.8 microM for the toxin-channel complex.  (+info)

Heterogeneity of the minimum functional unit of hemocyanins from the spider (Argiope bruennichii), the scorpion (Heterometrus sp.), and the horseshoe crab (Tachypleus tridentatus). (70/701)

The heterogeneity of arthropod hemocyanins was studied by polyacrylamide gel electrophoresis and immunochemical techniques. The spider (Argiope bruennichii), the scorpion (Heterometrus sp.), and the horseshoe crab (Tachypleus tridentatus) were found to have 4, 5, and 5 minimum functional units of hemocyanin, respectively, the apparent molecular weights of which were 79,000, 81000, and 80,000, respectively, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.  (+info)

The molecular structure of spider dragline silk: folding and orientation of the protein backbone. (71/701)

The design principles of spider dragline silk, nature's high-performance fiber, are still largely unknown, in particular for the noncrystalline glycine-rich domains, which form the bulk of the material. Here we apply two-dimensional solid-state NMR to determine the distribution of the backbone torsion angles (phi,psi) as well as the orientation of the polypeptide backbone toward the fiber at both the glycine and alanine residues. Instead of an "amorphous matrix," suggested earlier for the glycine-rich domains, these new data indicate that all domains in dragline silk have a preferred secondary structure and are strongly oriented, with the chains predominantly parallel to the fiber. As proposed previously, the alanine residues are predominantly found in a beta sheet conformation. The glycine residues are partly incorporated into the beta sheets and otherwise form helical structures with an approximate 3-fold symmetry.  (+info)

Energy metabolism of male and female tarantulas (Aphonopelma anax) during locomotion. (72/701)

We examined aerobic performance traits in male and female tarantulas (Aphonopelma anax). Reproductive fitness in these males relies heavily on locomotory searching to locate receptive females, which are fossorial and sedentary. Because of this dimorphism in life history, we predicted that selection in males would enhance their ability to sustain high levels of aerobic metabolism (compared with females) to support increased locomotory activity during the mating season. Rates of carbon dioxide production were measured in an enclosed variable-speed treadmill. Steady-state rates of carbon dioxide production increased linearly within the range of sustainable aerobic speeds for both males and females. Although there was substantial variation in physiological performance traits among individuals, there were no detectable intersexual differences in maximal rates of carbon dioxide production, maximal aerobic speed, minimum transport or factorial scope.  (+info)