Nascent chain dynamics and ribosome interactions within folded ribosome-nascent chain complexes observed by NMR spectroscopy.
Nascent chain dynamics and ribosome interactions within folded ribosome-nascent chain complexes observed by NMR spectroscopy.
复制标题
DOI：
10.1039/d1sc04313g
复制
发表时间：
2021-10-13
期刊：
影响因子：
8.4
通讯作者：
Christodoulou J
中科院分区：
文献类型：
--
作者：
Burridge C;Waudby CA;Włodarski T;Cassaignau AME;Cabrita LD;Christodoulou J
许多蛋白质的折叠可以在核糖体上的生物合成期间开始，并且可以由核糖体本身调节。这种扰动通常被认为是通过新生链和核糖体表面之间的相互作用介导的，但是尽管最近在表征未折叠状态与核糖体的相互作用方面取得了进展，以及它们对共翻译折叠的起始的影响，但新生链的折叠和未折叠状态之间的相互作用的完整定量分析尚未实现。在这里，我们应用溶液状态的NMR光谱测量横向质子弛豫速率的FLN 5细丝结构域的折叠核糖体新生链复合物中的甲基。我们观察到的新生链相对于孤立的域，这可以是相关的有效旋转相关时间的变化，使用测量的松弛和相互关联的松弛在孤立的域的松弛率大幅增加。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，并通过测量这些相互作用的变化，作为后续的FLN 6结构域出现，我们可以推断出它们对自由能景观的影响与共翻译折叠过程。核磁共振测量甲基弛豫在acetonally-arrested核糖体新生链复合物探测生物合成过程中出现的折叠新生多肽的动力学，并量化它们与核糖体表面的相互作用。
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. NMR measurements of methyl relaxation in translationally-arrested ribosome–nascent chain complexes probe the dynamics of folded nascent polypeptides emerging during biosynthesis and quantify their interaction with the ribosome surface.
登录
查看更多内容
影响因子：
16.8
作者：
Cabrita LD;Cassaignau AME;Launay HMM;Waudby CA;Wlodarski T;Camilloni C;Karyadi ME;Robertson AL;Wang X;Wentink AS;Goodsell L;Woolhead CA;Vendruscolo M;Dobson CM;Christodoulou J
通讯作者：
Christodoulou J
影响因子：
6.1
作者：
Anthis NJ;Clore GM
通讯作者：
Clore GM
DOI：
10.1126/science.1209740
发表时间：
2011-12-23
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kaiser CM;Goldman DH;Chodera JD;Tinoco I Jr;Bustamante C
通讯作者：
Bustamante C
DOI：
10.1073/pnas.74.4.1407
发表时间：
1977-01-01
影响因子：
11.1
作者：
LAVALETTE, D;AMAND, B;POCHON, F
通讯作者：
POCHON, F
影响因子：
15
作者：
Amero, Carlos;Schanda, Paul;Boisbouvier, Jerome
通讯作者：
Boisbouvier, Jerome