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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： Bin Wu Yan Zhu Jian Shi Jie Tao Yonghua Ji Lab of Neuropharmacology and Neurotoxicology Shanghai University Vascular Biology Research Centre Institute of Cardiovascular and Cell SciencesSt.George’sUniversity of LondonCranmer Terrace Central Laboratory Putuo HospitalShanghai University of Traditional Chinese Medicine

BmP02,a short-chain peptide with 28 residues from the venom of Chinese scorpion Buthus martensi Karsch,has been reported to inhibit the transient outward potassium currents(I) in rat ventricular muscle ***,it remains unclear whether BmP02 modulates the Kv4.2 channel,one of the main contributors to *** present study investigated the effects of BmP02 on Kv4.2 kinetics and its underlying molecular *** electrophysiological recordings showed that the inactivation of Kv4.2 expressed in HEK293 T cells was significantly delayed by BmP02 in a doseresponse manner with EC of～850 nM while the peak current,activation and voltage-dependent inactivation of Kv4.2 were not ***,the recovery from inactivation of Kv4.2 was accelerated and the deactivation was slowed after the application of *** site-directed mutagenesis combined with computational modelling identified that K347 and K353,located in the turret motif of the Kv4.2,and E4/E5,D20/D21 in BmP02 are key residues to interact with BmP02 through electrostatic *** findings not only reveal a novel interaction between Kv4.2 channel and its peptidyl modulator,but also provide valuable information for design of highly-selective Kv4.2 modulators.

关键词： scorpion toxin voltage-gated potassium channels electrophysiology interaction mode

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U-shaped dose-dependent effects of BmK AS,a unique scorpion polypeptide toxin on the voltage-gated sodium channels

U-shaped dose-dependent effects of BmK AS,a unique scorpion ...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Jie TAO~1 Mang-Mang ZHU~2 Miao TAN~2 Hong-Tian YANG~1 Yong-Hua JI~(1 *) 1 School of Life Sciences Shanghai University Shanghai 200444 China 2 Graduate School of Chinese Academy of Sciences Shanghai Institute of Physiology Shanghai Institute of Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

＜正＞Objective BmK AS,a scorpion polypeptide toxin,suggested to target the voltage-gated sodium channels （VGSCs）.However,the way of BmK AS acting on the VGSCs has yet to be *** We examined the electrophysiological effects of BmK AS in a wide dose range on the rat brain-type VGSC a-subunit,rNav1.2a,heterologously expressed in Xenopus oocytes as well as on the VGSCs endogenously expressed in the DRG neuroblastoma ND7- 23 cell ***（1） In the Xenopus oocytes,BmK AS depolarized the voltage dependence of activation and inactivation of rNav1.2a at 0.1 and 500 nM whereas hyperpolarized them at 1 nM.（2） While in the ND7-23 cells,BmK AS hyperpolarized the voltage dependence of activation and inactivation at 0.1,1 and 100 nM but not 10 *** AS also hyperpolarized the voltage dependence of recovery from inactivation at 0.1 and 100 nM,and slowed the recovery kinetics at all the concentrations,but the effects of 1 and 10 nM were relatively smaller than that of 0.1 and 100 nM.（3） The inactivation of VGSCs was potentiated by 10 nM BmK AS in both systems,whereas it was inhibited by 0.1 or 100 nM BmK AS in the Xenopus oocytes or ND7-23 cells,*** BmK AS could modulate the VGSCs in a unique Ushaped dose-dependent manner,which might be from an integration of the bidirectional effects via binding to two distinct receptor sites on the VGSCs.

关键词： U-shaped dose-dependence voltage-gated sodium channels polypeptide modulator scorpion toxin electrophysiology

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Purification,Characterization and ~1H NMR Resonance Assignment of an α-Like Neurotoxin BmK 16 from the Venom of Chinese scorpion Buthus martensii Karsch

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Chinese Journal of Chemistry 2003年第10期21卷 1378-1384页

作者：张乃霞吴宫王中华吴厚铭 State Key Laboratory of Bioorganic and Natural Products Chemistry

A natural scorpion toxin BmK 16 was purified for the first time from the venom of the Chinese scorpion Buthus martensii Karsch (BmK) by using combined gel filtration, ion exchange and reversed phase chromatography. The sequence of the N terminal 8 amino acid residues was determined by Edman degradation. Using the N terminal sequence as a tag, the database searching revealed a hit in the scorpion cDNA Bank. The sequence for N terminal 8 amino acid residues, molecular weight and amino acid compositions of BmK 16 were identical with the calculated values according to the first 64 residues' sequence of the precursor peptide alpha neurotoxin TX16 derived from the sequence of the cDNA AF156597 (EMBL). The sequence specific resonance assignment of BmK 16 was achieved and the intact sequence of BmK 16 was determined as followings: VRDAY IAKPH NCVYE CARNE YCNDL CTKNG AKSGY CQWVG KYGNG CWCKE LPDNV PIRVP GKCH. Furthermore, the results from the sequence homology analysis and the toxicity assays indicated that BmK 16 was an α like scorpion neurotoxin.

关键词： scorpion toxin Buthus martensii Karsch BmK 16,αlike scorpion neurotoxin amino acid sequence resonance assignment

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Recombinant scorpion insectotoxin AaIT kills specifically insect cells but not human cellss

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Cell Research 2002年第2期12卷 143-150页

作者： SHENG JIAN JI, FENG LIU, ER Qiu LI, Yu XIAN ZHUThe National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China 1The National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China 1The National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China 1The National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China 1The National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China

The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was confirmed by N-terminal peptide sequencing. Two groups of bioassays, artificial diet incorporation assay and contact insecticidal effect assay, were carried out separately to verify the toxicity of this recombinant toxin. At the end of a 24 h experimental period, more than 60% of the testing diamondback moth （Plutella xylostella） larvae were killed in both groups with LC50 value of 18.4 μM and 0.70 μM respectively. Cytotoxicity assay using cultured Sf9 insect cells and MCF-7 human cells demonstrated that the toxin AaIT had specific toxicity against insect cells but not human cells. Only 0.13 μM recombinant toxin was needed to kill 50% of cultured insect cells while as much as 1.3 μM toxin had absolutely no effect on human cells. Insect cells produced obvious intrusions from their plasma

关键词： scorpion toxin AaIT prokaryotic expression cytotoxicity.

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Structural basis of the SK Potassium Channel Controlling the Selectivity of Recognition by scorpion toxins

Structural basis of the SK Potassium Channel Controlling the...

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第十一届中国生物毒素研究及医药应用年会

作者： Jing Feng Youtian Hu Hong Yi Shijin Yin Song Han Jun Hu Zongyun Chen Weishan Yang Zhijian Cao Michel De Waard Jean-Marc Sabatier Wenxin Li 吴英亮 State Key Laboratory of Virology College of Life SciencesWuhan University Inserm U836 Grenoble Neuroscience InstituteLaboratory of Excellence 'Ion Channels Science and Therapeutics 'Universit Joseph FourierBtiment Edmond Safra38042 Grenoble Cedex 09France Inserm UMR1097 Aix-Marseille UniversiteParc Scientifique de Luminy13288 MarseilleFrance 武汉大学生命科学学院湖北省武汉市武汉大学生命科学学院

Potassium channel functions are often deciphered by using selective and potent scorpion *** these toxins,only a limited subset is capable to selectively block small conductance Ca-activated K(SK) *** sequence alignment,we found that SK channel turret residues are highly *** there are also some additional basic residues on the turret when compared with other potassium *** we presumed that there should be uniform structure bases of this selective SK channel *** this work, SK3 channel is exampled to demonstrate the key role of the electric charges of two conserved arginine residues(R485 and R489) from the SK3 channel outer vestibule in the selective recognition by SK3-blocking BmP05 ***,individually substituting these residues by histidyl or lysyl(maintaining partially or fully the positive electric charge),while decreasing the affinity of BmP05,still preserves its toxin-sensitivity profile(as evidenced by the lack of recognition by many other potassium channel-sensitive charybdotoxin(ChTX)).In contrast, when R485 or R489 residue of SK3 channel is mutated towards an acidic(E) or alcoholic(S) amino acid residue,the channel loses its sensitivity to BmP05 and becomes susceptible to a ’new’ blocking activity by *** these basic SK3 channel residues important for sensitivity,two acidic D492 and D518 residues,also located in the SK3 channel outer vestibule,were identified as being critical for toxin ***,molecular modeling data indicate the existence of a compact SK3 channel turret conformation,like a peptide screener, where the basic rings of R485 and R489 residues are stabilized by strong ionic interactions with D492 and D518 *** conclusion,the unique properties of two conserved arginine residues(spatial orientations and molecular interactions) of SK channel account for its toxin-sensitivity profile.

关键词： scorpion toxin recognition control conserved arginine residue SK3 channel outer vestibule electrostatic repulsion forces

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Inhibitory effects of recombinant neurotoxin BmK IM on seizures induced by pentylenetetrazol in Rats

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Chinese Medical Journal 2003年第12期116卷 1898-1903页

作者：何小华彭方章军建李文鑫曾宪春刘辉 Department of Biotechnology Department of Neurology

Objective To elucidate the inhibitory effects of recombinant Chinese scorpion neurotoxin BmK IM on seizures induced by pentylenetetrazol (PTZ) and the possible mechanism. Methods After purifying recombinant BmK IM from an E. coli cell line, its toxicity (both LD 50 and minimum lethal dose) on rats was determined. BmK IM was then microinjected into the CA3 region of the right hippocampus and its ability to inhibit the effects of an intraperitoneal injection of PTZ was assessed. The effects of BmK IM on the electrophysiological properties of isolated CA3 pyramidal neurons were then studied using whole-cell patch clamp techniques. Results BmK IM can significantly prolong the latent period of epileptic seizures, decrease the degree of seizures, and decrease the frequency of epileptiform discharges induced by PTZ. At the same time, 24h after injection of BmK IM into the hippocampal tissue, BmK IM significantly reduces the concentration of the neurotransmitter glutamate and alleviates PTZ-induced lesions in the hippocampus. Whole-cell patch clamp recordings indicate that BmK IM inhibits I Na of rat hippocampal neurons in a dose-dependent manner. BmK IM significantly shifts the activation curve of I Na in a positive direction, indicating that BmK IM enhances the threshold potential of I Na. Conclusions BmK IM has significant anti-epileptic properties, and may prove useful as a drug in the therapy of epilepsy. The inhibitory effects of BmK IM on seizures caused by pentylenetetrazol might depend on reductions in the release of presynaptic glutamate via the blocking of Na + channels.

关键词： scorpion toxin sodium channel epilepsy glutamate patch-clamp techniques hippocampus

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Novirus-mediated BmK CT expression suppresses growth and invasion of glioma

Novirus-mediated BmK CT expression suppresses growth and inv...

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泛环渤海地区九省市生物化学与分子生物学会——2011年学术交流会

作者： Jun Du Yue-Jun Fu Ai-Hua Liang Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education Institute of BiotechnologyShanxi University

BmK CT,one of the key toxins in scorpion Buthus martensii Karsch venom,can interact specifically with glioma cells as a chloride channel blocker and inhibit the invasion and migration of those *** this report,a recombinant adenovirus Ad-BmK CT was constructed and characterized by in vivo and in vitro *** results from cell proliferation assay with rat glioma(C6) cells showed that recombinant

关键词： scorpion toxin BmK CT Adenovirus-mediated,Gliomas.

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The recombinant GST-BmKCT fusion protein inhibits glioma proliferation and tumor metastasis

The recombinant GST-BmKCT fusion protein inhibits glioma pro...

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第九届中国生物毒素学术研讨会

作者： Shaozhong Fan~+,Zhengbo Sun~+,Dahe Jiang,Chao Dai,Yibao Ma,Zhenhuan Zhao,Hui Liu,Yingliang Wu, Zhijian Cao~(++),Wenxin Li~(++) State Key Laboratory of Virology,College of Life Sciences,Wuhan University,Wuhan,Hubei, 430072,PR China

Malignant gliomas are the most common primary brain tumors associated with significant morbidity and *** to target the tumor in situ,and inhibit tumor cell proliferation and invasion is the key for *** express a glioma-specific chloride ion channel that is sensitive to toxins including *** the current study,the inhibitory effect of BmKCT on glioma growth was observed in vivo using the glioma/SD rat ***, BmKCT prevented the metastasis of glioma cells in ***,biodistribution experiments with I-labeled or Cy5.5-conjugated BmKCT revealed that BmKCT selectively targeted the glioma in *** data suggest that BmKCT could be exploited as a potential therapeutic for glioma diagnosis and therapy.

关键词： scorpion toxin BmKCT Gliomas Proliferation Metastasis Targeting In vivo

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