ヤドクガエルAtelopus.zetekiの毒zetekitoxinの構造解析

箭毒蛙 Atelopus.zeteki 毒泽特毒素的结构分析

• 批准号: 12045210
• 负责人: 山下 まり
• 金额: $ 1.02万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas (A)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12045210/
• 关键词: ゼテキトキシン; カエル; 毒; サキシトキシン

中南米に生息するAtelopus(ヤセヤドクガエル)属のカエルの多くは、主に皮膚に水溶性毒を有する。1975年にコスタリカ産のAtelopus属のカエルからtetrodotoxinが同定されたが、zetekitoxinはすでに1969年に、Furman,Mosherによってパナマ産のA.zetekiに存在する新規の水溶性低分子毒として報告された。しかし、zetekitoxinは、その後、A.zetekiが保護条約下に置かれ、試料が入手出来なくなったことから、その構造は発見から約30年間未定であった。韓国科学技術院のY.H.Kim教授から規制前に精製された半精製品及び精製品のzetekitoxinを供与された。本研究は立体構造を含めたzetekitoxinの構造決定を行うことを目的とした。1.精製Biogel P-2,BioRex70,Hitachi gel3011Cを用いたHPLCで半精製品のzetekitoxinを精製し、純品を約300-400μg得た。2.性状FT-IR(ZnSe)1702,1561,1421,1338,1268cm^<-1>;HR-ESIMS[M+H]^+m/z553.1369(calcd for[C_<16>H_<25>N_8O_<12>S_1]^+m/z553.1313.[M-SO_3+H]^+m/z473.1725(calcd for[C_<16>H_<25>N_8O_9]^+m/z473.1744.この結果から、硫酸エステルの存在が示唆された。TLC(Silica gel 60)Pyr-EtOAc-AcOH-H_2O(15:7:3:6)Rf0.57(tetrodotoxin:0.50,saxitoxin:0.36).saxitoxinと同様に1%H_2O_2/H_2O(v/v)噴霧、加熱後にUV365nmで蛍光スポットとして検出された。3.NMRスペクトルと平面構造の推定NMRスペクトルは全て4%CD_3COOD/D_2O中、20℃か30℃で測定した。^1H-^1HCOSY,TNTOCSY,HSQC,HMBC,NOESY,NOE差,^<13>C NMRスペクトルの解析から、saxitoxin骨格を部分構造にウレタン、N-OHをもつ、非常に特異な新規構造を推定した。

In the middle and south of the country, there is a kind of water-soluble poison in the middle and south rice, which belongs to the genus Atelopus, which is mainly water-soluble. In 1975, there was a new regulation of water-soluble low molecular weight toxicants in water-soluble low molecular weight toxicants such as Atelopus, zetekitoxin, 1969, Furman,Mosher, and A.zeteki. After the purchase, zetekitoxin and post-treatment, the A.zeteki protection strip has not been scheduled for about 30 years, and the information has not been fixed for about 30 years. Professor Y.H.Kim of the National Institute of Science and Technology issued a standard for the production of semi-precision products and fine products zetekitoxin before the regulation. In this study, "three-dimensional" design includes "zetekitoxin", "decision", "action", "purpose" and "goal". 1. Biogel Pmuri 2 BioRex70 Hitachi gel3011C, "HPLC" semi-refined "zetekitoxin" essence, and "product" about 300-400 μ g. two。 Character FT-IR (ZnSe) 1761 lt;-1>;HR- 1421 1338, 1268cm ^ & lt;-1>;HR- EIMS [M + H] ^ + m/z553.1369 (calcd for [C _ & lt;16>H_<25>N_8O_<12>S_1] ^ + m/z553.1313. [M-SO_3+H] ^ + m/z473.1725 (calcd for [C _ & lt;16>H_<25>N_8O_9] ^ + m/z473.1744. The results showed that there was an instigation of sulfuric acid in the presence of sulfuric acid. TLC (Silica gel 60) Pyr-EtOAc-AcOH-H_2O Rf0.57 (tetrodotoxin:0.50,saxitoxin:0.36) .saxitoxin Rf0.57 (tetrodotoxin:0.50,saxitoxin:0.36) .saxitoxin is the same as the other 1%H_2O_2/H_2O (vmae), and after adding it, you can use the UV365nm light to get out of the box. The plane of 3.NMR has been constructed to determine the temperature of 20 ℃ and 30 ℃ in the full range of 4% CD _ 3COOD/D_2O, presumed by NMR. ^ 1H-^ 1HCOSYDON TNTOCSY HSQC department HMBC direction NOESYLONOE difference, ^ & lt;13>C NMR skeleton lattice analysis, saxitoxin bone lattice analysis, N-OH correction, special new rules, presumption, etc.

项目成果

Y.Shoi.,M.Yotsu-Yamashita et al.: "Electrospray Ionization Mass Spectrometry of Tetrodotoxin and its Analogs : Liquid Chromatography/Mass Spectrometry, Tandem Mass Spectrometry, and Liquid Chromatography/Tandem Mass Spectrometry"Anal, Biochem.. 290(1). 10

Y.Shoi.，M.Yotsu-Yamashita 等人：“河豚毒素及其类似物的电喷雾电离质谱法：液相色谱/质谱、串联质谱和液相色谱/串联质谱”Anal，Biochem.. 290（

E.J.Nunez-Vazquez,M,Yotsu-Yamashita et al.: "Toxicities and distribution of tetrodotoxin in the tissues on puffer fish found in the coast of the Baja California Peninsula, Mexico."Toxicon. 38. 729-734 (2000)

E.J.Nunez-Vazquez,M,Yotsu-Yamashita 等人：“墨西哥下加利福尼亚半岛海岸发现的河豚毒素的毒性和组织中的分布。”Toxicon。

M.Yotsu-Yamashita. et al.: "The levels of tetrodotoxin and its analogue 6-epitetrodotoxin in the red-spotted newts, Notophthalmus viridescens."Toxicon. (in press). (2001)

M.四山下。

H.Yotsu-Yamashita, et al.: "Binding Properties of ^3H-PbTx-3 and 3H-saxitoxin to Brain Membranes and to Skeletal Muscle Membranes of Puffer Fish Fugu pardalis, and the Primary Structure of a Voltage-Gated Na^+ Channel α-Subunit (fMNal) from Skeletal Muscl

H. Yotsu-Yamashita 等人：“^3H-PbTx-3 和 3H-石房蛤毒素与河豚河豚脑膜和骨骼肌膜的结合特性，以及电压门控 Na^+ 的一级结构来自骨骼肌的通道 α 亚基 (fMNal)