The Domains of Mammalian Base Excision Repair Enzyme N-Methylpurine-DNA Glycosylase

The Domains of Mammalian Base Excision Repair Enzyme N-Methylpurine-DNA Glycosylase
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哺乳动物碱基切除修复酶 N-甲基嘌呤-DNA 糖基化酶的结构域

DOI:
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发表时间:
1996
影响因子:
4.8
通讯作者:
S. Mitra
S. Mitra
中科院分区:
生物学2区
文献类型:
--
作者:
R. Roy;Amalendra Kumar;J. Lee;S. Mitra

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DNA中各种烷化碱基加合物的修复是通过N-甲基嘌呤-DNA糖基酶去除它们而启动的。31 kDa小鼠N-甲基嘌呤-DNA糖基酶是从333个氨基酸残基的野生型蛋白中缺失48个氨基酸残基而不失去活性的,分析了其是否存在在底物结合和催化中具有特定作用的蛋白酶抗性结构域。用胰酶增加蛋白分解,首先通过去除42个氨基末端残基产生29 kDa的多肽,然后产生具有明确的、不重叠的边界的8 kDa、6 kDa和13 kDa片段。8-kDa和13-kDa结构域分别包括氨基和羧基末端。根据DNA亲和层析和蛋白酶保护分析,6-kDa和13-kDa结构域似乎是非靶DNA结合所必需的,8-kDa结构域与其他两个结构域协同参与损伤碱基的识别。此外,化学交联研究表明,在底物DNA存在的情况下,8-kDa和6-kDa结构域发生构象变化,反映在保护蛋白降解和减少半胱氨酸残基对硫醇交换反应的可用性上。
Repair of a variety of alkylated base adducts in DNA is initiated by their removal by N-methylpurine-DNA glycosylase. The 31-kDa mouse N-methylpurine-DNA glycosylase, derived by deletion of 48 amino acid residues from the 333-residue wild type protein without loss of activity, was analyzed for the presence of protease-resistant domains with specific roles in substrate binding and catalysis. Increasing proteolysis with trypsin generated first a 29-kDa polypeptide by removal of 42 amino-terminal residues, followed by production of 8-, 6-, and 13-kDa fragments with defined, nonoverlapping boundaries. The 8- and 13-kDa domains include the amino and carboxyl termini, respectively. Based on DNA-affinity chromatography and the protease protection assay, it appears that the 6- and 13-kDa domains are necessary for nontarget DNA binding and that the 8-kDa domain, in cooperation with the other two domains, participates in recognition of damaged bases. Furthermore, chemical cross-linking studies indicated that, in the presence of substrate DNA, the 8- and 6-kDa domains undergo conformational changes reflected by both protection from proteolysis and reduced availability of cysteine residues for the thiol-exchange reaction.
使用单克隆抗体和有限的蛋白水解来阐明蛋白质的结构-功能关系。
DOI: 10.1002/9780470110560.ch4
发表时间: 1991
期刊: Methods of biochemical analysis
影响因子: --
作者:
Wilson,JE
通讯作者: Wilson,JE
DOI: 10.1126/science.6351251
发表时间: 1983-01-01
期刊: SCIENCE
影响因子: 56.9
作者:
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DOI: --
发表时间: 1994
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Kumar,A
DOI: 10.1016/s0021-9258(18)48324-x
发表时间: 1987-05
期刊: The Journal of biological chemistry
影响因子: --
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通讯作者: S. Randall;Ramon Eritjaj;Bruce E. Kaplanj;John PetruskaS;M. Goodman
DOI: 10.1073/pnas.90.19.8901
发表时间: 1993-10-01
影响因子: 11.1
作者:
BESSHO, T;ROY, R;MITRA, S
通讯作者: MITRA, S