两种虎纹捕鸟蛛昆虫毒素的分离纯化及生物学活性鉴定

结合离子交换和反相高效液相色谱从虎纹捕鸟蛛粗毒分离纯化到 2种虎纹捕鸟蛛毒素 ,命名为虎纹捕鸟蛛毒素 Ⅶ和虎纹捕鸟蛛毒素 Ⅷ .经质谱测定这 2种毒素的分子量分别为 3981 0 2和 4 171 12 .氨基酸序列分析发现 ,这 2种虎纹捕鸟蛛毒素同源性非常高 ,只有 6个残基位点的不同 .虎纹捕鸟蛛毒素 Ⅶ和虎纹捕鸟蛛毒素 Ⅷ的生物学功能相似 ,都能对蝗虫起到麻痹作用 ;对小鼠的中枢神经作用高剂量能使小鼠产生致死 ,但低剂量虎纹捕鸟蛛毒素 Ⅷ能使小鼠产生惊厥反应 ,而低剂量虎纹捕鸟蛛毒素 Ⅶ不能使小鼠产生惊厥反应 ;这 2种毒素都能阻断小鼠离体膈神经膈肌的神经肌肉传递 ,且与虎纹捕鸟蛛毒素 I混合后都具协同作用     

虎纹捕鸟蛛毒素-III及其天然突变体的纯化与鉴定(英文)

虎纹捕鸟蛛毒素 III及其天然突变体是从虎纹捕鸟蛛粗毒中分离得到的两个毒素多肽。虎纹捕鸟蛛毒素 III含 33个氨基酸残基 ,其中包含 6个半胱氨酸残基 ;而其天然突变体只比虎纹捕鸟蛛毒素 III少了C端的色氨酸残基。MALDI TOF质谱测得虎纹捕鸟蛛毒素 III及其天然突变体的分子量分别为 385 3.35和 36 6 7.4 0。通过比较其理论分子量和质谱测定的分子量表明两个多肽的 6个半胱氨酸残基分别形成了三对二硫键。虎纹捕鸟蛛毒素 III与从同一种蜘蛛分离得到的凝集素 I具有 70 .5 %的序列相似性。生物学活性实验表明 ,虎纹捕鸟蛛毒素 III具有使美洲蜚蠊可逆的致瘫作用 ,其半有效剂量 (ED50 )为 (1 92 .95±1 2 0 .84 ) μg/g (P =0 .95 ) ,而且能加强由电刺激引起的大鼠输精管收缩 ;而其天然突变体却不具有上述生物学活性 ,表明C端色氨酸残基为虎纹捕鸟蛛毒素 III生物学活性相关残基 ;同时虎纹捕鸟蛛毒素 III及其天然突变体都不具有类似于凝集素 I对红细胞的凝集活性 ,表明虎纹捕鸟蛛毒素 III和凝集素 I两者氨基酸序列中不同氨基酸残基对于决定两者的生物学活性有着重要的作用     

虎纹捕鸟蛛实验种群越冬及其耐饥力的观察

在长沙地区,虎纹捕鸟蛛幼蛛和成蛛的入蛰时间分别为11月中旬和11月底至12月上旬;出蛰时间分别为翌年3月下旬和3月中旬,冬眠期蜘蛛的成活率主要由土温高低决定.虎纹捕鸟蛛具有较强的耐饥力.     

温度和食物对虎纹捕鸟蛛发育和繁殖力影响的研究

研究不同温度和食物对虎纹捕鸟蛛生长发育和繁殖力的影响,结果表明,虎纹捕鸟蛛在２４℃－２６℃变温范围,饲以人工混合饲料溶液时生长发育最快,在这个温度范围外饲以单一食物为最低。最有利于虎纹捕鸟蛛繁殖的因子水平组合为２４℃－２６℃,食物为人工混合饲料溶液。     

虎纹捕鸟蛛毒素Ⅴ的二硫键定位分析

虎纹捕鸟蛛毒素Ⅴ是从虎纹捕鸟蛛毒液中分离得到的一种昆虫毒素.它含有35个氨基酸残基,其中6个半胱氨酸形成三对二硫键.首先采用多酶将天然的肽链裂解后,通过MALDI-TOF质谱分析酶解肽段,推断出1对二硫键位于Cys9-Cys21,然后利用改进的部分还原分步测序法,确定虎纹捕鸟蛛毒素Ⅴ的另外2对二硫键的配对方式为Cys2-Cys16和Cys15-Cys28.因此,虎纹捕鸟蛛毒素Ⅴ的3对二硫键分别以Cys2-Cys16,Cys9-Cys21,Cys15-Cys28（即1-4、2-5和3-6）的方式配对.     

我国捕鸟蛛两种雄蛛的描述(蛛形纲,蜘蛛目)

中国捕鸟蛛科2)过去记述5种.除虎纹捕鸟蛛Ornithoctonus huwena外,其它4种均为单性描述.经补充采集,发现了广西近捕鸟蛛Plesiophrictus guangxiensis Yin et Tan,2000和海南捕鸟蛛Selenocosmia hainana Liang,Peng,Wang et Chen,1999的雄性个体.补充描述如下.标本保存在湖南师范大学生命科学学院,文中量度单位均为mm.     

虎纹捕鸟蛛发育起点温度和有效积温研究

实验设置5个温度、3个湿度。对虎纹捕鸟蛛卵袋和幼蛛的发育速率进行测定。结果表明：温度是影响虎纹捕鸟蛛发育的重要因子。湿度对卵历期、幼蛛历期有显著影响。运用优选法计算虎纹捕鸟蛛发育起点温度和有效积温。所得卵和全世代的发育起点温度T0分别为14．3℃。7．0℃。有效积温K分别为712．66日度和6161．80日度。     

温湿度对虎纹捕鸟蛛繁殖力影响的研究

研究不同温湿度对虎纹捕鸟蛛繁殖力的影响,结果表明：虎纹捕鸟蛛繁殖力在26℃恒温和RH80％的因子水平组合中较大。在24－26℃变温和RH80％的组合中为最大。     

虎纹捕鸟蛛生物学特性的观察

虎纹捕鸟蛛（Ornithoctonus huwenna)个体大,毒性强,有强烈的攻击性,蛛毒含量大,采毒易,是进行蛛毒,蛛蛋白研究的理想试验材料,研究在饲养条件下虎纹捕鸟蛛的生活习性,捕食特点,雌蛛产卵,护卵习性,卵的孵化,蜕皮及幼蛛的发育,以及耐饥饿,耐干旱能力。     

蜘蛛寄生革螨1新属2新种(蜱螨亚纲:巨刺螨科)

报道巨刺螨科Macronyssidae 1新属2新种:广西蛛刺螨 Arachnyssus guang.riensis gen.et sp. nov.和虎纹蛛刺螨 Arachnyssus huwenae gen.et sp. nov., 均寄生于虎纹捕鸟蛛 Ormthoctonus huwena Wang et al., 1993。     

虎纹捕鸟蛛体表扫描电镜观察

对虎纹捕岛蛛(Ornithoctonus husoena)的眼、生殖球、颚叶、听毛、触毛、琴形器、跗节器、发音器、幼蛛及成蛛的纺绩器和螯肢等结构进行了扫描电镜观察。     

The cDNA and genomic DNA organization of a novel toxin SHT-I from spider Ornithoctonus huwena

Spider venoms are complex mixtures of neurotoxicpeptides, proteins and low molecular mass organicmolecules. Their neurotoxic activity is due to the interac-tion of the venom components with cellular receptors, inparticular ion channels. Spider venoms have…     

Genomic organization and cloning of novel genes encoding toxin-like peptides of three superfamilies from the spider Orinithoctonus huwena

The bird spider Ornithoctonus huwena is one of the most venomous spiders in China. Its venom is a mixture of various compounds with diverse bioactivities. Ninety proteins and 47 peptides have been identified, and 67 cDNA sequences encoding different toxin precursors have been cloned. However, the genomic DNA of them is seldom reported. To characterize the genomic DNA structure of huwentoxins, the genomic DNA encoding toxins of three superfamilies were cloned by using sequence specific or partially degenerate primers based on their cDNA sequences. An unexpected finding was that the intron was lacking in the genomic sequences of three superfamilies. The genomic DNA information has predictive value for better understanding the relationship of spider toxin evolution. In addition, we have cloned and analyzed 19 novel genes encoding toxin-like precursors by using the genomic DNA of the spider O. huwena.     

温度对蜘蛛繁殖力和实验种群的影响

从我国常见10科18种蜘蛛的温度实验数据中,找出温度对蜘蛛繁殖和实验种群增长影响的一般规律和不同种类蜘蛛繁殖的最佳温度。根据实验可以看出：大多数蜂蛛在1－35℃温区内均可产卵和孵化;由于蜘蛛种类不同,雌蛛产卵袋数最多、卵袋内含卵量最高、产卵间隔时间最短、产卵率最高、单雌产卵量最多、孵化率最高和种群增长指数最大的最适温度也表现出差异;大多数种类雌蛛产卵袋数最多的温区是25－32℃;产卵间隔最短的温区是30－37℃;产卵率最高的温区是25－32℃;孵化率最高的温区在25℃左右;种群数量增长最多的温区是25－29℃。温度与蜘蛛繁殖力的关系可以用曲线方程和数学模型表示。     

虎纹捕鸟蛛神经细胞急性分离培养及其电压门控通道膜片钳

探索了虎纹捕鸟蛛(Ornithoctonus huwena)食道下神经细胞急性分离培养条件,并利用全细胞膜片钳技术对虎纹捕鸟蛛食道下神经细胞电压门控性钠、钾和钙通道的基本电生理学特性进行了研究.适合虎纹捕鸟蛛神经细胞离体培养的培养基为(g/L):葡萄糖0.7,果糖0.4,琥珀酸0.06,咪唑0.06,L-1513.7,Hepes 2.38,酵母粉2.8,乳白蛋白2.5,青霉素200 IU/ml,链霉素200 mg/ml,小牛血清15%;pH 6.8.该培养基非常适合虎纹捕鸟蛛神经节神经细胞离体培养,细胞在温度(27±2)℃的培养箱中培养2～4h,培养的细胞数目多、结构完整、贴壁效果好,细胞近似汤勺形,有一个长的单极突起,大部分细胞在10～30μm之间.全细胞模式下可以记录到钠、钾和钙三种电压门控离子通道电流.钙电流为高电压激活电流,该电流能够被NiCl2完全抑制;钾电流为瞬时钾电流和延迟整流钾电流,这两类钾电流分别被细胞外液中的4-氨基吡啶和氯化四乙胺所阻断;钠电流为TTX敏感型电流.     

The cross channel activities of spider neurotoxin huwentoxin-I on rat dorsal root ganglion neurons

In this paper, we investigated the action of huwentoxin-I (HWTX-I) purified from the venom of the Chinese bird spider Ornithoctonus huwena on Ca(2+), Na(+) channels of adult rat dorsal root ganglion (DRG) neurons. The results showed that huwentoxin-I could reduce the peak currents of N-type Ca(2+) channels (IC(50) approximately 100 nM) and TTX-S Na(+) channels (IC(50) approximately 55 nM), whereas no effect was detected on TTX-R Na(+) channels. The comparative studies indicated that the selectivity of HWTX-I on Ca(2+) channels was higher that of MVIIA and approximately the same as that of GVIA. HWTX-I is the first discovered toxin with the cross channel activities from the spider O. huwena venom similar to micro O-conotoxins MrVIA and MrVIB.     

Proteomic and peptidomic characterization of the venom from the Chinese bird spider, Ornithoctonus huwena Wang

The bird spider Ornithoctonus huwena Wang is a very venomous spider in China. Several compounds with different types of biological activities have been identified previously from the venom of this spider. In this study, we have performed a proteomic and peptidomic analysis of the venom. The venom was preseparated into two parts: the venom proteins with molecular weight (MW) higher than 10,000 and the venom peptides with MW lower than 10 000. Using one-dimensional gel electrophoresis (1-DE), two-dimensional gel electrophoresis (2-DE), and mass spectrometry, 90 proteins were identified, including some important enzymes, binding proteins, and some proteins with significant biological functions. For venom peptides, a combination of cation-exchange and reversed-phase chromatography was employed. More than 100 components were detected by mass spectrometry, and 47 peptides were sequenced by Edman degradation. The peptides display structural and pharmacological diversity and share little sequence similarity with peptides from other animal venoms, which indicates the venom of O. huwena Wang is unique. The venom peptides can be classified into several superfamilies. Also it is revealed that gene duplication and focal hypermutation have taken place during the evolution of the spider toxins.     

Comparative pharmacology and cloning of two novel arachnid sodium channels: Exploring the adaptive insensitivity of scorpion to its toxins

Scorpion toxins have been found lacking effect on Na(+) current of its own sodium channel, whereas the molecular mechanism remains mystery. In this study, the binding affinity of pharmacologically distinct scorpion toxins was found much weaker to scorpion (Buthus martensii) nerve synaptosomes than to spider (Ornithoctonus huwena) ones. The sodium channel cDNA from these two species were further cloned. The deduced proteins contain 1871 and 1987 amino acids respectively. Several key amino acid substitutions, i.e., A1610V, I1611L and S1617K, are found in IVS3-S4 constituting receptor site-3, and for receptor site-4, two residues (Leu-Pro) are inserted near IIS4 of scorpion sodium channel.     

虎纹捕鸟蛛毒素虎纹毒素-III对美洲蜚蠊神经细胞电压门控离子通道的影响

HWTX-III是从中国虎纹捕鸟蛛Ornithoctonus huwena粗毒中分离纯化到的一种昆虫神经多肽。通过应用全细胞膜片钳技术研究了HWTX-III对美洲蜚蠊Periplaneta americana神经细胞电压门控离子通道的影响。发现HWTX-III特异性地抑制美洲蜚蠊背侧不成对中间(dorsal unpaired median, DUM）神经细胞的电压门控钠通道(IC50≈1.106 μmol/L),而对电压门控钾通道没有明显的影响。HWTX-III通过一种新型的不同于其他蜘蛛毒素的机制抑制昆虫电压门控钠通道,它不影响通道的激活与失活动力学,也不明显地漂移稳态失活曲线。HWTX-III对昆虫神经细胞电压门控钠通道的特异性与新型作用机制为研究电压门控钠通道分子结构的多样性以及开发新的安全的杀虫剂提供有用的工具。