Domain organization and functional analysis of Thermus thermophilus MutS protein
Domain organization and functional analysis of Thermus thermophilus MutS protein
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DOI：
10.1093/nar/26.18.4153
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发表时间：
1998-09-15
影响因子：
14.9
通讯作者：
Kuramitsu, S
中科院分区：
文献类型：
--
作者：
Tachiki, H;Kato, R;Kuramitsu, S
MutS蛋白结合DNA并特异性识别错配或小环出的异源双链DNA。为了阐明其结构与功能的关系，通过变性实验和有限蛋白酶解研究了嗜热栖热菌MutS蛋白的结构域。前者表明嗜热嗜热菌MutS至少由三个结构域组成，其稳定性估计为12.3、22.9和30.7 kcal/mol;后者表明其由四个结构域组成：A1（N-末端至残基130）、A2（131-274）、B（275-570）和C（571-C-末端）。凝胶阻滞试验表明，嗜热嗜热厌氧杆菌MutS与双链DNA（ds）非特异性相互作用，但不与单链DNA相互作用。在蛋白水解片段中，B结构域与dsDNA结合。在这些结果的基础上，我们提出了结构域的组织嗜热嗜热菌MutS和这些结构域的推定作用。
MutS protein binds to DNA and specifically recognizes mismatched or small looped out heteroduplex DNA. In order to elucidate its structure-function relationships, the domain structure of Thermus thermophilus MutS protein was studied by performing denaturation experiments and limited proteolysis. The former suggested that T.thermophilus MutS consists of at least three domains with estimated stabilities of 12.3, 22.9 and 30.7 kcal/mol and the latter revealed that it consists of four domains: A1 (N-terminus to residue 130), A2 (131-274), B (275-570) and C (571 to C-terminus). A gel retardation assay indicated that T.thermophilus MutS interacts non-specifically with double-stranded (ds), but not single-stranded DNA. Among the proteolytic fragments, the B domain bound to dsDNA. On the basis of these results we have proposed the domain organization of T.thermophilus MutS and putative roles of these domains.