期刊论文

Wang, X.C.

中文

生物工程学报

1000-3061

2000

16

4

490-494

10.3321/j.issn:1000-3061.2000.04.017

国家高技术研究发展计划项目!（ 10 3 13 0 1 0 6）；

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虎纹捕鸟蛛毒素 Ⅰ （Huwentoxin Ⅰ ,HWTX Ⅰ ）是从虎纹捕鸟蛛 （Selenocosmiahuwena）的粗毒中分离出的一种多肽类神经毒素。为了探明该毒素分子中唯一的Arg残基与其生物学活性的关系 ,运用固相多肽合成技术和Fmoc化学直接构建了Ala取代HWTX Ⅰ第 2 0位Arg（R2 0 ）的突变体R2 0A HWTX Ⅰ ;将合成的突变体置于含谷胱甘肽的缓冲体系中氧化复性后用反相和特殊设计的离子交换HPLC纯化 ,并对之进行氨基酸组成、Edman降解与质谱分析。活性测定结果表明 ,HWTX Ⅰ分子中的R2 0被A取代后 ,活性下降了 92 % ,提示R2 0是与活性密切相关的重要残基。

Huwentoxin-I (HWTX-I) is a polypeptide neurotoxin purified from the venom of the spider Selenocosmia huwena. R20A-HWTX-I, a mutant of HWTX-I in which the Arg was replaced by Ala, was synthesized on solid support by using Fmoc chemistry. The synthetic mutant was oxidatively renatured in glutathione-containing buffer, and then isolated by reversed phase and specially designed ion-exchange HPLC. The chemical structure of R20A-HWTX-I was confirmed by amino acid analysis, Edman degradation and MALDI-TOF mass analysis. Physiological experiment showed that the replacement of R20 by a decreased the bi...