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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Single-particle EM reveals extensive conformational variability of the Ltn1 E3 ligase
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Single-particle EM reveals extensive conformational variability of the Ltn1 E3 ligase

机译:单粒子EM揭示Ltn1 E3连接酶的广泛构象变异

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摘要

Ltn1 is a 180-kDa E3 ubiquitin ligase that associates with ribosomes and marks certain aberrant, translationally arrested nascent poly-peptide chains for proteasomal degradation. In addition to its evolutionarily conserved large size, Ltn1 is characterized by the presence of a conserved N terminus, HEAT/ARM repeats predicted to comprise the majority of the protein, and a C-terminal catalytic RING domain, although the protein's exact structure is unknown. We used numerous single-particle EM strategies to characterize Ltni's structure based on negative stain and vitreous ice data. Two-dimensional classifications and subsequent 3D reconstructions of electron density maps show that Ltn1 has an elongated form and presents a continuum of conformational states about two flexible hinge regions, whereas its overall architecture is reminiscent of multisubunit cullin-RING ubiquitin ligase complexes. We propose a model of Ltn1 function based on its conformational variability and flexibility that describes how these features may play a role in cotranslational protein quality control.
机译:Ltn1是一个180 kDa的E3泛素连接酶,与核糖体缔合并标记某些异常的,翻译中止的新生多肽链,用于蛋白酶体降解。 Ltn1除了进化上保守的大分子外,其特征还包括保守的N末端,HEAT / ARM重复序列预计占蛋白质的大部分以及C端催化RING域,尽管该蛋白质的确切结构尚不清楚。我们使用了许多单粒子EM策略来根据负染色和玻璃冰数据表征Ltni的结构。二维分类和随后的电子密度图3D重建显示Ltn1具有细长的形式,并在两个柔性铰链区域周围呈现连续的构象状态,而其总体结构让人联想到多亚基cullin-RING泛素连接酶复合物。我们基于其构象变异性和灵活性提出了一个Ltn1功能模型,该模型描述了这些功能如何在共翻译蛋白质量控制中发挥作用。

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