蛇毒锯鳞蝰素基因Leul4－Lys15－Glu16的定点突变

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	蛇毒锯鳞蝰素基因Leul4－Lys15－Glu16的定点突变; Site-directed mutagenesis of the gene of Echistatin Leu14-Lys15-Glu16
	李洪超 ; 李雄彪 ; 胡美浩
刊名	遗传学报
	1996
关键词	蛇毒锯鳞蝰素定点突变抑制血小板凝集活性 PCR
英文摘要	本研究的目的是利用蛋白质工程定点突变的方法，在蛇毒锯鳞蝰素基因分子上增加另一个保守序列ＲＧＤ（１４位精氨酸残基，１５位甘氨酸残基，１６位天冬氨酸残基），以其增加该分子的生物活性，并探讨蛋白质一级结构，空间结构和功能的关系。在质粒ｐＪＣ２６４的基础上，利用ＰＣＲ定点突变方法，对蛇毒锯鳞蝰素基因Ｌｅｕ１４－Ｌｙｓ１５－Ｇｌｕ１６进行定点突变，使相应的ＤＮＡ片段变成表达Ａｒｇ１４－Ｇｌｙ１５－Ａｓｐ１６的核苷酸顺序，经酶切和ＤＮＡ测序鉴定正确。ＣＮＢｒ裂解后，用反相ＨＰＬＣ分析，分离制备突变体蛇毒锯鳞蝰素，制备的突变体蛇毒锯鳞蝰素的Ｎ－末端１０个氨基酸残基与天然蛇毒锯鳞蝰素的相同。在人的富含血小板血...; In order to probe relationship of inhibition activity of platelet aggregation and RGD conformation beneficial to binding in Echistatin, we used the site-directed mutation technique to install another RGD sequence into one of irregular loops retaining a degree of conformational flexibility and substituting Leu-14, Lys-15, Glu-16 of (Leu-28) Echistatin. The mutant (Arg-14, Gly-15, Asp-16, Leu-28) Echistatin did not lose its inhibition activity of platelet aggregation; however, it showed at least as high activity as (Leu-28) Echistatin, or even a little higher than (Leu-28) Echistatin. This suggested that both RGD sequences inserted in one loop with a degree of conformational flexibility. The original RGD (Arg-24, Gly-25, Asp-26) motif projecting significantly from the surface of the scaffold or core might contribute synergistically to the function of inhibiting platelet aggregation induced by 10 mumol/ L ADP (final concentration). These results are useful in the elucidation of the relationship of structure and function of Echistatin-like disintegrins and GPIIb/IIIa-like integrins.; PubMed; 中文核心期刊要目总览(PKU); 中国科学引文数据库(CSCD); 0; 02; 163-168; 23
语种	中文
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/45640]
专题	生命科学学院
推荐引用方式 GB/T 7714	李洪超,李雄彪,胡美浩. 蛇毒锯鳞蝰素基因Leul4－Lys15－Glu16的定点突变, Site-directed mutagenesis of the gene of Echistatin Leu14-Lys15-Glu16[J]. 遗传学报,1996.
APA	李洪超,李雄彪,&胡美浩.(1996).蛇毒锯鳞蝰素基因Leul4－Lys15－Glu16的定点突变.遗传学报.
MLA	李洪超,et al."蛇毒锯鳞蝰素基因Leul4－Lys15－Glu16的定点突变".遗传学报 (1996).