细菌 tRNA(Sec) 与丝氨酰-tRNA 合成酶复合物的结晶和初步 X 射线晶体学分析。

IF 0.9 4区生物学

Acta Crystallographica Section F-structural Biology and Crystallization Communications Pub Date : 2012-06-01 Epub Date: 2012-05-23 DOI:10.1107/S1744309112016004

Yuzuru Itoh, Shun Ichi Sekine, Shigeyuki Yokoyama

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引用次数: 0

摘要

硒半胱氨酸（Sec）通过翻译结合到蛋白质中，以响应 UGA 密码子。硒半胱氨酸（Sec）的特异性 tRNA（tRNA(Sec)）首先通过丝氨酸-tRNA 合成酶（SerRS）与丝氨酸连接。为了阐明细菌 tRNA（Sec）的三级结构及其与 SerRS 的特异性相互作用，将来自 Aquifex aeolicus 的细菌 tRNA（Sec）与来自 Methanopyrus kandleri 的古生物 SerRS 作为异源复合物进行了结晶。虽然 tRNA(Sec) 与野生型 SerRS 复合物晶体的 X 射线衍射效果很差（分辨率为 5.7 Å），但通过引入针对晶体-包装界面的点突变，分辨率得到了提高。重原子标记也有助于提高分辨率。从 K(2)Pt(CN)(4)-soaked 晶体中获得了用于相测定的 3.2 Å 分辨率衍射数据集。

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Crystallization and preliminary X-ray crystallographic analysis of bacterial tRNA(Sec) in complex with seryl-tRNA synthetase.

Selenocysteine (Sec) is translationally incorporated into proteins in response to the UGA codon. The tRNA specific to Sec (tRNA(Sec)) is first ligated with serine by seryl-tRNA synthetase (SerRS). To elucidate the tertiary structure of bacterial tRNA(Sec) and its specific interaction with SerRS, the bacterial tRNA(Sec) from Aquifex aeolicus was crystallized as the heterologous complex with the archaeal SerRS from Methanopyrus kandleri. Although X-ray diffraction by crystals of tRNA(Sec) in complex with wild-type SerRS was rather poor (to 5.7 Å resolution), the resolution was improved by introducing point mutations targeting the crystal-packing interface. Heavy-atom labelling also contributed to resolution improvement. A 3.2 Å resolution diffraction data set for phase determination was obtained from a K(2)Pt(CN)(4)-soaked crystal.

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来源期刊

Acta Crystallographica Section F-structural Biology and Crystallization Communications 生物-晶体学

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审稿时长

2-4 weeks

期刊介绍： Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.