Nano-channels in the spider fang for the transport of Zn ions to cross-link His-rich proteins pre-deposited in the cuticle matrix

We identify the presence of multiple vascular channels within the spider fang. These channels seem to serve the transport of zinc to the tip of the fang to cross-link the protein matrix by binding to histidine residues. According to amino acid and elemental analysis of fangs extracted shortly after ecdysis, His-rich proteins are deposited before Zn is incorporated into the cuticle. Microscopic and spectroscopic investigations in the electron microscope and synchrotron radiation experiments suggest that Zn ions are transported through these channels in a liable (yet unidentified) form, and then form stable complexes upon His binding. The resulting cross-linking through the Zn–His complexes is conferring hardness to the fang. Our observations of nano-channels serving the Zn-transport within the His-rich protein matrix of the fibre reinforced spider fang may also support recent bio-inspired attempts to design artificial polymeric vascular materials for self-healing and in-situ curing.     

虎纹捕鸟蛛毒素-Ⅰ突变体A1Y-HWTX-Ⅰ的固相合成和生物学活性测定

用芴甲氧羰基 (Fmoc)固相多肽合成的方法在自制自动蛋白质化学工作站上合成了用酪氨酸 (Y)替代虎纹捕鸟蛛毒素 - (HWTX- )第一位丙氨酸 (A1 )的突变体 A1 Y- HWTX- .合成的突变体用 Edman降解和电喷雾质谱法进行鉴定 .活性分析结果证明 ,合成的 A1 Y- HWTX- 在含有谷胱甘肽的缓冲体系中氧化折叠后显示出与天然 HWTX- 完全相同的生物学活性 ,提示 Y替代 HWTX- 的 A1后并不明显影响 HWTX- 的活性部位和空间构象 ;A1与 HWTX- 生物学活性无关 .此外 ,将 Y引入 HWTX- 分子有助于利用碘标记方法研究 HWTX- 的作用机制     

蜘蛛丝的分子结构与力学性能研究

蜘蛛丝尤其是蜘蛛大囊状腺产生的拖丝,具有独特的机械性能,是自然界颇具应用潜力的生物材料。现代分子生物学技术使蜘蛛丝蛋白基因得以克隆,通过高分子物理化学手段方法的利用,有利于揭示蜘蛛丝蛋白质序列、分子结构、以及分子结构和力学性能之间的关系。对不同种类蜘蛛丝蛋白的深入研究,将为基因工程方法人工合成并改造蜘蛛丝成为可能。     

Molecular cloning,bioinformatics analysis and functional characterization of HWTX-XI toxin superfamily from the spider Ornithoctonus huwena

Spider venom contains a very valuable repertoire of natural resources to discover novel components for molecular diversity analyses and therapeutic applications. In this study, HWTX-XI toxins from the spider venom glands of Ornithoctonus huwena which are Kunitz-type toxins (KTTs) and were directly cloned, analyzed and functionally characterized. To date, the HWTX-XI superfamily consists of 38 members deduced from 121 high-quality expressed sequence tags, which is the largest spider KTT superfamily with significant molecular diversity mainly resulted from cDNA tandem repeats as well as focal hypermutation. Among them, HW11c40 and HW11c50 may be intermediate variants between native Kunitz toxins and sub-Kunitz toxins based on evolutionary analyses. In order to elucidate their biological activities, recombinant HW11c4, HW11c24, HW11c27 and HW11c39 were successfully expressed, further purified and functionally characterized. Both HW11c4 and HW11c27 display inhibitory activities against trypsin, chymotrypsin and kallikrein. Moreover, HW11c4 is also an inhibitor relatively specific for Kv1.1 channels. HW11c24 and HW11c39 are found to be inactive on chymotrysin, trypsin, kallikrein, thrombin and ion channels. These findings provide molecular evidence for toxin diversification of the HWTX-XI superfamily and useful molecular templates of serine protease inhibitors and ion channel blockers for the development of potentially clinical applications.     

三种蜘蛛丝蛋白组成分析

本文应用高压液相色谱（ＨＰＬＣ）法分析了岳麓山的大腹园蛛Ａｒａｎｅｕｓ ｖｅｎｔｒｉｃｏｓｕｓ（Ｃ．Ｋｏｃｈ,１８７８）,机敏漏头蛛Ａｇｅｌｅｎａ ｄｉｆｆｉｃｌｉｓ （Ｆｏｘ,１９３７）,白额巨蟹蛛Ｈｅｔｅｒｏｐｏｄａ ｖｅｎａｔｏｒｉａ （Ｌｉｎｎａｅｕｓ,１７５７）的丝蛋白的氨基酸组成,以ＳＤＳ－ＰＡＧＥ法测定了大腹园蛛不同丝腺体的未成丝的可溶性丝蛋白的分子量。实验结果表明蛛丝蛋白中占优势的     

Screening of Necrotizing Spiders in Korea Using Nonradioactive Sphingomyelinase Assay(II) Web-building Spiders

Spider bites cause a range of symptoms from simple swellings to disfiguring necrotic lesions, and occasionally death. While spider bites are not a major medical problem in Korea, it would be of great value to know which species of spiders pose a threat to human health. A middle molecular weight protein, sphingomyelinase D, has been identified in the venom of the brown recluse spider and strong evidence suggests that they have a major role in spider bite necrosis. For the identification of necrotizing species, we have investigated using recently developed non‐radioactive assay of sphingomyelinase for rapidly screening the necrotizing venoms. Here, we demonstrate the fetal toxicity of total 65 species (24 genera, 9 families) of the web‐building spiders among 622 identified spider species in Korea. It has been revealed that four species of the orb‐weaving spider, Araneus ventricosus (family Araneidae, 0.3509), Dipoena castrata (0.2413, family Theridiidae), Argiope minuta (0.1836, family Araneidae), and Paracoelotes spinivulva (0.1760, family Amaurobiidae) have relatively strong activities among themselves. However, comparing to that of the brown recluse spider, Loxosceles recluse (1.814) in North America, the necrotizing shpingomyelinase activities of these Korean web‐building spiders are still very low. Based on our results, we may thus conclude that there would be little possibilities in South Korea to create serious medical problems caused by necrotizing arachnidism.     

Inheritance of sex ratio in the Kanzawa spider mite,Tetranychus kanzawai Kishida

Population Ecology - The frequency distributions in sex ratios of offspring from 4 fieldTetranychus kanzawai populations were examined. No significant difference was observed between average sex...     

Central nervous projections of mechanoreceptors in the spider Cupiennius salei Keys.

Summary The basic organization of sensory projections in the suboesophageal central nervous system of a spider (Cupiennius salei Keys.) was analyzed with anterograde cobalt fills and a modified Golgi rapid method. The projections of three lyriform slit sense organs and of tactile hairs located proximally on the legs are described and related to central nerve tracts. There are five main longitudinal sensory tracts in the central region of the suboesophageal nervous mass arranged one above the other. Whereas the three dorsal ones contain fibers from the lyriform organs, the two ventral ones contain axons from the hair receptors. Axons from all three lyriform organs have typical shapes and widely arborizing ipsilateral intersegmental branches and a few contralateral ones. The terminal branches of the afferent projections from identical lyriform organs on each leg form characteristic longitudinal pathways, typical of each organ: U-shaped, O-shaped, or two parallel bundles. The terminations of the hair sensilla are ipsilateral and intersegmental. Two large bilaterally arranged longitudinal sensory association tracts receive inputs from all legs including the dense arborizations from tactile hairs, lyriform organs, and other sense organs. These tracts may serve as important integrating neuropils of the suboesophageal central nervous system.     

Stochastic prey arrivals and crab spider giving-up times: simulations of spider performance using two simple “rules of thumb”

Summary We compared the patch-choice performances of an ambush predator, the crab spider Misumena vatia (Thomisidae) hunting on common milkweed Asclepias syriaca (Asclepiadaceae) umbles, with two stochastic rule-of-thumb simulation models: one that employed a threshold giving-up time and one that assumed a fixed probability of moving. Adult female Misumena were placed on milkweed plants with three umbels, each with markedly different numbers of flower-seeking prey. Using a variety of visitation regimes derived from observed visitation patterns of insect prey, we found that decreases in among-umbel variance in visitation rates or increases in overall mean visitation rates reduced the clarity of the optimum (the difference in the yield obtained as foraging behavior changes), both locally and globally. Yield profiles from both models were extremely flat or jagged over a wide range of prey visitation regimes; thus, differences between optimal and next-best strategies differed only modestly over large parts of the foraging landscape. Although optimal yields from fixed probability simulations were one-third to one-half those obtained from threshold simulations, spiders appear to depart umbels in accordance with the fixed probability rule.