Nuclear Magnetic Resonance Studies on Huwentoxin-Ⅺ from the Chinese Bird Spider Ornithoctonus huwena:^(15)N Labeling and Sequence-specific ~1H,^(15)N Nuclear Magnetic Resonance Assignments  

作　　者：Kuan PENG Ying LIN Song-Ping LIANG 

机构地区：[1]College of Life Science, Peking University, Beijing 100871, China [2]College of Life Science, Hunan Normal University, Changsha 410081, China

出　　处：《Acta Biochimica et Biophysica Sinica》2006年第7期457-466,共10页生物化学与生物物理学报（英文版）

基　　金：This work was supported by the grants from the National Natural Science Foundation of China(No.30170193 and No.30430170)

摘　　要：Huwentoxin-XI purified from the Chinese bird spider Ornithoctonus huwena is a toxin with both antiprotease activity and potassium channel blocking activity. To determine its solution structure, huwentoxin-XI was expressed in a yeast eukaryotic expression system and studied by NMR. The ^15N labeling strategy was used to facilitate the process of resonance assignments. The nearly complete sequence-specific assignments of proton and nitrogen resonances were obtained by analyzing a series of two-dimensional （2D） and three-dimensional （3D） spectra, including DQF-COSY, TOCSY, NOESY, ^15N-^1H HSQC, ^15N-^1H HNHA, ^15N-^1H HNHB, ^15N-^1H TOCSY-HSQC and ^15N-^1H NOESY-HSQC spectra. Secondary structure analysis of huwentoxin-XI showed that it mainly contains an N-terminal 310-helix from Thr3 to Arg5 and a C-terminal α-helix from Gln45 to Cys52, plus a triple-stranded antiparallel β-sheet of Glu18-Asn23, Thr26-Ile31 and Asn40-Lys41. These studies provide a solid basis for the final structure determination of huwentoxin-XI.Huwentoxin-XI purified from the Chinese bird spider Ornithoctonus huwena is a toxin with both antiprotease activity and potassium channel blocking activity. To determine its solution structure, huwentoxin-XI was expressed in a yeast eukaryotic expression system and studied by NMR. The ^15N labeling strategy was used to facilitate the process of resonance assignments. The nearly complete sequence-specific assignments of proton and nitrogen resonances were obtained by analyzing a series of two-dimensional （2D） and three-dimensional （3D） spectra, including DQF-COSY, TOCSY, NOESY, ^15N-^1H HSQC, ^15N-^1H HNHA, ^15N-^1H HNHB, ^15N-^1H TOCSY-HSQC and ^15N-^1H NOESY-HSQC spectra. Secondary structure analysis of huwentoxin-XI showed that it mainly contains an N-terminal 310-helix from Thr3 to Arg5 and a C-terminal α-helix from Gln45 to Cys52, plus a triple-stranded antiparallel β-sheet of Glu18-Asn23, Thr26-Ile31 and Asn40-Lys41. These studies provide a solid basis for the final structure determination of huwentoxin-XI.

关 键 词：huwentoxin-XI TOXIN nuclear magnetic resonance ^15N labeling sequence-specific assignment 

分 类 号：Q959.7[生物学—动物学]