嗜热栖热菌MutS蛋白的结构域组织与功能分析
Domain organization and functional analysis of Thermus thermophilus MutS protein.
作者信息
Tachiki H, Kato R, Masui R, Hasegawa K, Itakura H, Fukuyama K, Kuramitsu S
机构信息
Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan.
出版信息
Nucleic Acids Res. 1998 Sep 15;26(18):4153-9. doi: 10.1093/nar/26.18.4153.
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.
MutS蛋白与DNA结合，并特异性识别错配或小的环状异源双链DNA。为了阐明其结构-功能关系，通过进行变性实验和有限蛋白酶解研究了嗜热栖热菌MutS蛋白的结构域结构。前者表明嗜热栖热菌MutS至少由三个结构域组成，估计稳定性分别为12.3、22.9和30.7千卡/摩尔，后者表明它由四个结构域组成：A1（N端至第130位残基）、A2（131-274）、B（275-570）和C（571至C端）。凝胶阻滞试验表明，嗜热栖热菌MutS与双链（ds）而非单链DNA非特异性相互作用。在蛋白水解片段中，B结构域与dsDNA结合。基于这些结果，我们提出了嗜热栖热菌MutS的结构域组织以及这些结构域的假定作用。
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