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通过定点诱变产生核糖核酸酶抗体酶

Creation of a ribonuclease abzyme through site-directed mutagenesis.

作者信息

Fletcher M C, Kuderova A, Cygler M, Lee J S

机构信息

Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

出版信息

Nat Biotechnol. 1998 Nov;16(11):1065-7. doi: 10.1038/3519.

DOI:10.1038/3519
PMID:9831037
Abstract

The development of abzymes (antibody/enzymes) is one method of creating reagents with novel catalytic activity. To date, most abzymes have been obtained by immunization with transition state analogs. We have chosen to start with an existing antibody and convert it into an enzyme by the addition of catalytic residues to the binding site. We have introduced a histidine residue into antibody Jel 103 and converted it into an abzyme that cleaves poly(rI) with a kinetic efficiency of about 100 M(-1) sec(-1).

摘要

抗体酶(抗体/酶)的开发是一种创造具有新型催化活性试剂的方法。迄今为止,大多数抗体酶是通过用过渡态类似物进行免疫获得的。我们选择从一种现有的抗体开始,通过在结合位点添加催化残基将其转化为酶。我们已将一个组氨酸残基引入抗体Jel 103,并将其转化为一种抗体酶,该抗体酶切割聚(rI)的动力学效率约为100 M⁻¹ s⁻¹。

相似文献

1
Creation of a ribonuclease abzyme through site-directed mutagenesis.通过定点诱变产生核糖核酸酶抗体酶
Nat Biotechnol. 1998 Nov;16(11):1065-7. doi: 10.1038/3519.
2
Site-directed mutagenesis of active site contact residues in a hydrolytic abzyme: evidence for an essential histidine involved in transition state stabilization.水解抗体中活性位点接触残基的定点诱变:参与过渡态稳定的必需组氨酸的证据。
J Mol Biol. 1997 Apr 18;267(5):1247-57. doi: 10.1006/jmbi.1997.0938.
3
In vitro abzyme evolution to optimize antibody recognition for catalysis.体外抗体酶进化以优化抗体对催化作用的识别。
Nat Biotechnol. 2001 Jun;19(6):563-7. doi: 10.1038/89320.
4
Directed evolution governed by controlling the molecular recognition between an abzyme and its haptenic transition-state analog.通过控制抗体酶与其半抗原过渡态类似物之间的分子识别来进行定向进化。
J Immunol Methods. 2004 Nov;294(1-2):1-14. doi: 10.1016/j.jim.2004.06.018.
5
Positional ordering of reacting groups contributes significantly to the efficiency of proton transfer at an antibody active site.反应基团的位置排序对抗体活性位点处质子转移的效率有显著贡献。
J Am Chem Soc. 2005 Feb 2;127(4):1307-12. doi: 10.1021/ja044647l.
6
Molecular mechanisms of improvement of hydrolytic antibody 6D9 by site-directed mutagenesis.通过定点诱变改善水解抗体6D9的分子机制
J Biochem. 2006 Oct;140(4):509-15. doi: 10.1093/jb/mvj179. Epub 2006 Aug 18.
7
Enzyme mimicry by the antiidiotypic antibody approach.通过抗独特型抗体方法进行酶模拟。
Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13526-31. doi: 10.1073/pnas.200360497.
8
Probing the importance of second sphere residues in an esterolytic antibody by phage display.通过噬菌体展示探究酯解抗体中第二球域残基的重要性。
J Mol Biol. 1998 Dec 11;284(4):1083-94. doi: 10.1006/jmbi.1998.2234.
9
Catalytic antibodies (abzymes) induced by stable transition-state analogs.由稳定过渡态类似物诱导产生的催化抗体(抗体酶)。
Biochemistry (Mosc). 2000 Nov;65(11):1233-44.
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Cloning and expression of a single-chain catalytic antibody that acts as a glutathione peroxidase mimic with high catalytic efficiency.一种具有高催化效率、可模拟谷胱甘肽过氧化物酶功能的单链催化抗体的克隆与表达。
Biochem J. 2001 Oct 15;359(Pt 2):369-74. doi: 10.1042/0264-6021:3590369.

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Antibody manufacture in transgenic animals and comparisons with other systems.转基因动物中的抗体生产及其与其他系统的比较。
Curr Opin Biotechnol. 2002 Dec;13(6):625-9. doi: 10.1016/s0958-1669(02)00362-2.