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Transcriptomic analysis of the Latrodectus tredecimguttatus spider venomous gland using Expressed sequence tags

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Transcriptomic analysis of the Latrodectus tredecimguttatus ...

第九届中国生物毒素学术研讨会

作者： Duan zhigui Wang Meichi Wang Xianchun liang songping~* Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China,Hunan Normal University,Changsha Hunan 410081,China

＜正＞Black widow spider is one of the most poisonous spiders in the *** to now, there have been few systematic analyses of the spider venom components,and the mechanism of action of the venom has not been completely ...

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Synthesis and characterization of huwentoxin-Ⅳ,a neurotoxin inhibiting central neuronal sodium channels

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Synthesis and characterization of huwentoxin-Ⅳ,a neurotoxin...

Activating Na＿v1.9 Recovers Pain Responses in Na＿v1.7-related CIP as Revealed by a Spider Toxin

第八届中国生物毒素学术研讨会

作者： Xuan Luo# Yucheng Xiao# Fang Kuang Meichun Deng Meichi Wang songping liang~* Life Sciences College,Hunan Normal University,Changsha,Hunan 410081,P.R.China

Our previous work demonstrated that huwentoxin-Ⅳ was an inhibitor cystine knot peptide from Chinese tarantula Ornithoctonus huwena venom that blocked tetrodotoxin-sensitive voltage-gated sodium channels from mammalian sensory ***,the actions of the neurotoxin on central neuronal sodium channels remain *** this study,we chemically synthesized native huwentoxin-Ⅳ and found that sodium channel isoforms from rat hippocampus neurons were also sensitive to native and synthetic toxins,but the toxin binding affinity (IC50 .4 μM) was 12-fold lower than to peripheral *** blockade by huwentoxin-Ⅳ could be reversed by strong depolarization due to the dissociation of toxin-channel complex as observed for receptor site 3 ***,small unilamellar vesicles binding assays showed that in contrast to ProTx-Ⅱ from the tarantula Thrixopelma pruriens,huwentoxin-Ⅳ lacked the ability to partition into the negatively charged and neutral phospholipid bilayer of artificial *** findings indicated that huwentoxin-Ⅳ was a sodium channel antagonist preferentially targeting peripheral isoforms via a mechanism quite different from ProTx-Ⅱ.

关键词： spider toxin sodium channel whole-cell recording hippocampus

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Activating Na＿v1.9 Recovers Pain Responses in Na＿v1.7-rela...

第十四届生物毒素毒理学术大会暨第一届生物毒素——从生存适应到转化医学专题学术会议

作者： Xi Zhou Tingbin Ma Luyao Yang Shuijiao Peng Lulu Li Zhouquan Wang Zhen Xiao Qingfeng Zhang Li Wang Yazhou Huang Minzhi Chen songping liang Xianwei Zhang Jing Yu Liu Zhonghua Liu The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development College of Life SciencesHunan Normal University Key Laboratory of Molecular Biophysics of the Ministry of Education College of Life Science and TechnologyHuazhong University of Science and Technology(HUST) Department of Anesthesiology Tongji Hospital of HUST

In peripheral nociceptor,Na1.7,Na1.8 and Na1.9 are critical for pain ***,loss-of-function of Na1.7 leads to congenital insensitivity to pain(CIP) in humans,which causes individuals to loss the "warning system" and thus suffer from recurrent tissue damage and serious *** far,the therapeutic agents for treating the Nav1.7-related CEP are still in its *** we identified and characterized a spider peptide toxin called HpTx1 that reverses alganesthesia in Na1.7-KO mice by enhancing the excitability of DRG *** elucidate this process,we conformed that HpTxl inhibits Na 1.7(IC=0.51±0.12 μM),but it activates Nav 1.9(EC=0.47±0.08 μM) and has no effects on ***,HpTx1 reduces dorsal root ganglion(DRG) neurons excitability and produces analgesia in Na1.9-KO mice,and losses its effectiveness in Na1.8-KO mice These data indicated the pain-inducing effect by HpTxl should be mediated by Na1.9 ***,we found that HpTxl interacts with DIV s3 b-s4 of Na1.9 and DII s3 b-s4 of Na1.7,respectively,which explains its opposite effects on the two *** together,our findings suggest that Na1.9 agonists such as HpTx1 may be potential for the development of drugs to treat Na1.7-related CIP,and propose that Na1.7,Na1.8 and Na1.9 are interrelated in pain signaling.

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Huwentoxin-I,a multifunctional toxin that preferentially targets insect voltage-gated sodium channels

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Huwentoxin-I,a multifunctional toxin that preferentially tar...

第九届中国生物毒素学术研讨会

作者： Meichi Wang,Mingqiang Rong,Yongqun Zhang,Zhonghua Liu,songping liang~* Key Laboratory of Protein Chemistry and Developmental Biology of the Ministry of Education,the College of Life Science,Hunan Normal University,Changsha,PR China,410081

＜正＞Huwentoxin-Ⅰis a 33-amino acid peptide isolated from the crude venom of Ornithoctonus *** synthesized Huwentoxin-Ⅰand demonstrated that the sodium channel inhibition of synthetic Huwentoxin-Ⅰ（sHuwentoxin-Ⅰ... 详细信息

关键词： tetrodotoxin-sensitive voltage-gated sodium channel dorsal root ganglion dorsal unpaired median hippocampal neuron

Arg20突变对敬钊缨毛蛛毒素-V钠通道活性的影响

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生物工程学报 2008年第7期24卷 1228-1232页

作者：曾雄智邓梅春皮建辉全妙华王贤纯梁宋平湖南师范大学蛋白质化学与发育生物学教育部重点实验室长沙410081 怀化学院生物工程系怀化418008

敬钊缨毛蛛毒素-V(Jingzhaotoxin-V,JZTX-V)是从敬钊缨毛蛛粗毒中纯化到的一种新型河豚毒素不敏感型钠通道抑制剂,为了深入研究该毒素的结构与功能关系,应用芴甲氧羰基(Fmoc)固相多肽化学合成方法合成了用丙氨酸(Ala)替代JZTX-V第20位... 详细信息

敬钊缨毛蛛毒素-V(Jingzhaotoxin-V,JZTX-V)是从敬钊缨毛蛛粗毒中纯化到的一种新型河豚毒素不敏感型钠通道抑制剂,为了深入研究该毒素的结构与功能关系,应用芴甲氧羰基(Fmoc)固相多肽化学合成方法合成了用丙氨酸(Ala)替代JZTX-V第20位精氨酸残基的突变体R20A-JZTX-V,合成线性多肽经反相高效液相色谱分离纯化后进行谷胱甘肽氧化复性。复性产物分别用基质辅助激光解析飞行时间质谱(MALDI-TOF/TOF MS)进行分子量的鉴定,用膜片钳电生理方法进行电压门控钠通道抑制活性分析。研究结果表明,Arg20被Ala取代后,R20A-JZTX-V对大鼠背根神经节细胞(DRG)膜上表达的河豚毒素敏感型(TTX-S)钠通道的抑制活性与天然JZTX-V相当,提示Arg20与JZTX-V对TTX-S钠通道的抑制活性无关或关系不大;而R20A-JZTX-V对TTX-R钠通道的抑制活性却比天然JZTX-V下降了约18.3倍,说明Arg20是与JZTX-V对河豚毒素不敏感型(TTX-R)钠通道抑制活性相关的关键活性残基之一,推测R20A-JZTX-V活性降低的原因是用Ala替代Arg20后改变了JZTX-V与TTX-R型钠通道的作用位点。

关键词：敬钊缨毛蛛毒素-V固相多肽合成突变体氧化复性膜片钳

衍生性聚偏二氟乙烯膜用于蛋白质及肽的固相序列测定

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湖南医学 1999年第6期16卷 406-408页

作者：吴学玲陈平梁宋平湖南医学高等专科学校生化教研室长沙410006 湖南师范大学生命科学学院长沙410081

【目的】研究衍生聚偏二氟乙烯(PVDF) 膜用于蛋白质及肽的固相序列测定的可行性。【方法】在PVDF 膜上引入对苯二异硫氰酸基(DITC) 制成能与蛋白质及肽共价偶联的膜载体。用这种膜载体与蛋白质或肽样品偶联后,进行Edman 降解及氨基酸顺... 详细信息

【目的】研究衍生聚偏二氟乙烯(PVDF) 膜用于蛋白质及肽的固相序列测定的可行性。【方法】在PVDF 膜上引入对苯二异硫氰酸基(DITC) 制成能与蛋白质及肽共价偶联的膜载体。用这种膜载体与蛋白质或肽样品偶联后,进行Edman 降解及氨基酸顺序分析。【结果】该膜载体化学性能稳定,偶联操作简单,结果准确,与蛋白质及肽的偶联率为78 .19 % ,Edman降解的重复回收率为90 .27 % ,所测氨基酸序列与已知序列完全相同。【结论】衍生PVDF

关键词：氨基酸序列膜,人工乙烯类聚合物

Large-scale identification and analysis of peptide toxins from the tarantula Ornithoctonus hainana venom using a venomic strategy

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Large-scale identification and analysis of peptide toxins fr...

第九届中国生物毒素学术研讨会

作者： Xing Tang~#,Yongqun Zhang~#,Weijun Hu,Dehong Xu,Huai Tao,Xiaoxu Yang,Yan Li,songping liang~* The Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education,College of Life Sciences,Hunan Normal University,Changsha 410081, China

＜正＞The tarantula Ornithoctonus hainana is a very venomous spider spread in the hilly areas of Hainan province in southern *** venoms contain a variety of toxic components with different pharmacological *** the present study,we used a venomic strategy for high-throughput identification of tarantula-venom peptides from *** strategy include three different approaches:（i） transcriptomics,i.e., EST based cloning and PCR-based cloning plus DNA sequencing（ii） peptidomics, i.e.,off-line MDLC-MS plus peptide sequencing（direct Edman sequencing and bottom-up sequencing）（iii） genomics,i.e.,genomic DNA cloning plus DNA *** 420 non-redundant peptides were detected by mass spectrometry, 272 peptide precursor were characterized from cDNA or genomic DNA,and 192 full or partial sequences peptides were sequenced by multiple *** the basis of the precursor sequences,all peptide toxins from the tarantula *** venom can be classified into 11 superfamilies（and related families）.Phylogenetics analyses revealed that gene duplication and focal hypermutation may be responsible for the enormous molecular and pharmacological diversity in spider *** current work is the initial overview for the study of tarantulas-venom in parallel transcriptomic, peptidomic,genomic ***,this work will also provide an effective high-throughput methodology for venom-peptide identification from other spider species,especially tarantulas species.

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虎纹捕鸟蜘蛛毒素对全脑缺血大鼠海马神经元细胞形态及bax、bcl-2 mRNA和蛋白表达的影响

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中国运动医学杂志 2006年第6期25卷 678-683页

作者：毛海峰陈嘉勤梁宋平王一蓉刘仁仪湖南师范大学体育学院长沙410012 湖南师范大学生命科学学院

目的:观察蛛网膜下腔注射虎纹捕鸟蜘蛛毒素(HWTX-I)对全脑缺血再灌注大鼠海马神经细胞线粒体和CA1区锥体细胞形态、凋亡相关因子bax和bcl-2 mRNA及蛋白表达的影响。方法:SD大鼠随机分为假手术组、生理盐水组及HWTX-I组,采用改良的Pulsin... 详细信息

目的:观察蛛网膜下腔注射虎纹捕鸟蜘蛛毒素(HWTX-I)对全脑缺血再灌注大鼠海马神经细胞线粒体和CA1区锥体细胞形态、凋亡相关因子bax和bcl-2 mRNA及蛋白表达的影响。方法:SD大鼠随机分为假手术组、生理盐水组及HWTX-I组,采用改良的Pulsinelli四血管阻断全脑缺血损伤结合蛛网膜下腔置管术模型,对大鼠海马神经细胞线粒体进行超微结构及Nissl染色形态学观察,免疫组织化学法检测Bax、Bcl-2蛋白表达,RT-PCR法检测baxb、cl-2 mRNA表达。结果:(1)HWTX-I能够使全脑缺血大鼠海马神经细胞线粒体基本维持正常形态。(2)HWTX-I能稳定全脑缺血大鼠海马CA1区锥体细胞分布。(3)HWTX-I能够下调全脑缺血再灌注大鼠海马组织bax mRNA及蛋白表达,上调bcl-2 mRNA及蛋白表达。结论:蛛网膜下腔注射HWTX-I对全脑缺血再灌注所致的大鼠海马神经元损伤有明显的保护作用。

关键词：虎纹捕乌蜘蛛毒素(HWTX-I) 全脑缺血再灌注线粒体 bcl-2 bax

豆科植物籽实中的昆虫消化道α－淀粉酶抑制剂的研究（Ⅱ）

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生物技术 1995年第2期5卷 29-31页

作者：王贤纯梁宋平湖南师范大学生物系

对５个不同品种菜豆（ＰｈａｓｅｏｌｕｓｖｕｌｇａｌｒｉｕｓＬ．）中的蜚蠊（Ｐｅｒｉｐｌａｎｅｔａ）消化道α－淀粉酶抑制剂作了比较研究．发现它们都能在酸性ｐＨ条件下有效地发挥抑制作用，且最适抑制ｐＨ均在５．５０左右；但... 详细信息

对５个不同品种菜豆（ＰｈａｓｅｏｌｕｓｖｕｌｇａｌｒｉｕｓＬ．）中的蜚蠊（Ｐｅｒｉｐｌａｎｅｔａ）消化道α－淀粉酶抑制剂作了比较研究．发现它们都能在酸性ｐＨ条件下有效地发挥抑制作用，且最适抑制ｐＨ均在５．５０左右；但它们的热稳定性、最大抑制活性及发挥最大抑制活性所需的预温时间等不尽相同。

关键词：蛋白质类抑制剂 α-淀粉酶菜豆蜚蠊豆科