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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Structural insights into the adaptation of proliferating cell nuclear antigen (PCNA) from haloferax volcanii to a high-salt environment
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Structural insights into the adaptation of proliferating cell nuclear antigen (PCNA) from haloferax volcanii to a high-salt environment

机译：结构适应的见解增殖细胞核抗原(PCNA)haloferax volcanii高盐环境

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摘要

The sliding clamp proliferating cell nuclear antigen (PCNA) plays vital roles in many aspects of DNA replication and repair in eukaryotic cells and in archaea. Realising the full potential of archaea as a model for PCNA function requires a combination of biochemical and genetic approaches. In order to provide a platform for subsequent reverse genetic analysis, PCNA from the halophilic archaeon Haloferax volcanii was subjected to crystallographic analysis. The gene was cloned and expressed in Escherichia coli and the protein was purified by affinity chromatography and crystallized by the vapour-diffusion technique. The structure was determined by molecular replacement and refined at 3.5 ? resolution to a final R factor of 23.7% (R free = 25%). PCNA from H. volcanii was found to be homotrimeric and to resemble other homotrimeric PCNA clamps but with several differences that appear to be associated with adaptation of the protein to the high intracellular salt concentrations found in H. volcanii cells.
机译：滑动夹增殖细胞的核抗原(PCNA)在许多方面扮演着重要的角色真核细胞的DNA复制和修复古生菌。古生菌增殖细胞核抗原函数需要一个模型生化和基因的组合方法。随后的反向遗传分析,PCNA的的嗜盐的archaeon Haloferax volcanii进行晶体分析。克隆和表达在大肠杆菌和蛋白质被亲和纯化色谱法和结晶vapour-diffusion技术。由分子置换和雅致在3.5吗?免费(R = 25%)。homotrimeric和其他类似homotrimeric PCNA夹但与几个似乎相关的差异适应高的蛋白质细胞内的盐浓度在H。volcanii细胞。

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