Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy,Chemical Science

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Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy
Chemical Science ( IF 7.6 ) Pub Date : 2021-09-09 , DOI: 10.1039/d1sc04313g
Charles Burridge ₁ , Christopher A Waudby ₁ , Tomasz Włodarski ₁ , Anaïs M E Cassaignau ₁ , Lisa D Cabrita ₁ , John Christodoulou ₁

Affiliation

The folding of many proteins can begin during biosynthesis on the ribosome and can be modulated by the ribosome itself. Such perturbations are generally believed to be mediated through interactions between the nascent chain and the ribosome surface, but despite recent progress in characterising interactions of unfolded states with the ribosome, and their impact on the initiation of co-translational folding, a complete quantitative analysis of interactions across both folded and unfolded states of a nascent chain has yet to be realised. Here we apply solution-state NMR spectroscopy to measure transverse proton relaxation rates for methyl groups in folded ribosome–nascent chain complexes of the FLN5 filamin domain. We observe substantial increases in relaxation rates for the nascent chain relative to the isolated domain, which can be related to changes in effective rotational correlation times using measurements of relaxation and cross-correlated relaxation in the isolated domain. Using this approach, we can identify interactions between the nascent chain and the ribosome surface, driven predominantly by electrostatics, and by measuring the change in these interactions as the subsequent FLN6 domain emerges, we may deduce their impact on the free energy landscapes associated with the co-translational folding process.

中文翻译：

通过核磁共振波谱观察折叠核糖体-新生链复合物中的新生链动力学和核糖体相互作用

许多蛋白质的折叠可以在核糖体上的生物合成过程中开始，并且可以由核糖体本身调节。人们普遍认为这种扰动是通过新生链和核糖体表面之间的相互作用介导的，但尽管最近在表征未折叠状态与核糖体的相互作用及其对共翻译折叠启动的影响方面取得了进展，但对新生链的折叠和展开状态之间的相互作用尚未实现。在这里，我们应用溶液态核磁共振波谱来测量 FLN5 细丝蛋白结构域的折叠核糖体-新生链复合物中甲基的横向质子弛豫率。我们观察到相对于隔离域，新生链的弛豫率大幅增加，这可能与使用隔离域中的弛豫和互相关弛豫测量的有效旋转相关时间的变化有关。使用这种方法，我们可以识别新生链和核糖体表面之间的相互作用，主要由静电驱动，并且通过测量这些相互作用随着后续 FLN6 域出现而发生的变化，我们可以推断它们对与 FLN6 域相关的自由能景观的影响。共平移折叠过程。

更新日期：2021-09-15

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