Identification, characterization, and purification of DNA glycosylase/AP lyases by reductive crosslinking to 2'-deoxyribooligonucleotides containing specific base lesions.
Identification, characterization, and purification of DNA glycosylase/AP lyases by reductive crosslinking to 2'-deoxyribooligonucleotides containing specific base lesions.
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通过与含有特定碱基损伤的 2-脱氧核糖寡核苷酸进行还原交联来鉴定、表征和纯化 DNA 糖基酶/AP 裂解酶。
DOI：
10.1006/meth.2000.1059
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Cunningham,RP
中科院分区：
文献类型：
--
作者：
Chheda,AD;Teebor,GW;Cunningham,RP
本文描述了一种还原胺化交联法，它有助于鉴定和鉴定一类与碱基切除修复有关的DNA修复酶--DNA糖基酶/AP裂解酶。这种交联技术已被用于鉴定粗提物和部分纯化的酶制剂中的酶，分离蛋白质进行测序，并确定该酶家族成员的反应机理。席夫碱基酶-底物的化学还原为稳定的胺，导致位于2‘-脱氧寡核苷酸内的底物损伤与修复酶之间形成不可逆共价键。该复合体可以通过凝胶电泳法检测到，也可以通过氨基酸序列分析来分离和分析。
This paper describes a reductive amination crosslinking protocol that facilitates identification and characterization of a class of DNA repair enzymes, DNA glycosylase/AP lyases, which are involved in base excision repair. This crosslinking technique has been used to identify enzymes in crude extracts and in partially purified enzyme preparations, to isolate proteins for sequencing, and to confirm the reaction mechanism of members of this enzyme family. Chemical reduction of the Schiff's base enzyme–substrate intermediate to a stable amine results in the formation of an irreversible covalent bond between the substrate lesion situated within a 2′-deoxyoligonucleotide and the repair enzyme. This complex can be detected by gel electrophoresis and can also be isolated and analyzed by amino acid sequencing.
影响因子：
14.9
作者：
Z. Hatahet;A. Purmal;S. Wallace
通讯作者：
Z. Hatahet;A. Purmal;S. Wallace
硼氢化钠与肌肉磷酸化酶1的反应
DOI：
--
发表时间：
1958
期刊：
影响因子：
--
作者：
E. Fischer;A. B. Kent;Eloise. Snyder;E. Krebs
通讯作者：
E. Krebs
影响因子：
2.9
作者：
Hilbert, TP;Boorstein, RJ;Teebor, GW
通讯作者：
Teebor, GW