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在出生时绘制蛋白质-蛋白质相互作用:对核糖体新生球蛋白复合物的单颗粒冷-电磁分析

Meranda M Masse¹, Rachel B Hutchinson¹, Christopher E Morgan²

¹Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.

ACS central science

|March 4, 2024

概括

这项研究揭示了阿波米oglobin (apoMb) 在共翻译折叠过程中如何与核糖体相互作用. 新生链与核糖体蛋白 (r-蛋白) 和RNA相互作用,特别是L23,在核糖体上.

科学领域:

分子生物学分子生物学
结构生物学结构生物学
生物化学生物化学

背景情况:

与核糖体组件的核糖体结合新生链 (RNC) 相互作用对于共翻译蛋白质折叠至关重要.
关于RNC-核糖体接触的先前研究主要使用C-终端停滞序列,在没有这些动机的情况下了解相互作用时留下了一个空白.
关于新生链和核糖体外表面上的核糖体蛋白质 (r-蛋白质) 之间的相互作用的信息很少.

研究的目的:

为了研究与核糖体结合的球蛋白 (apoMb) 的结构特征及其与核糖体的相互作用.
探索在核糖体出口道内及其外表面的新生链-核糖体接触,特别是在没有强烈的滞后序列的情况下.
阐明特定核糖体组件,包括r-蛋白和rRNA在新生的链折叠和相互作用中的作用.

主要方法:

化学交叉链接用于识别新生链和核糖体之间的近位残留物.
单粒子冷电子显微镜 (cryo-EM) 用于高分辨率的结构确定.
光异构性衰变以评估新生链的动态及其相互作用.

主要成果:

在核糖体退出道核心内,apoMb相互作用类似于之前报告的发现.
当RNC进入道前廊时,它表现出增强的动态,并与核糖体RNA (rRNA) 和L23r蛋白相互作用.
在核糖体的外表面,RNCs主要与L23r蛋白上保存的非极性补丁相互作用.
ApoMb的N端区域是动态和紧的,缺乏与核糖体的直接接触.
在转位过程中,阿波米oglobin 通过其 C-终端区域与核糖体相互作用.

结论:

氨基球蛋白的配译折叠涉及与核糖体的动态相互作用,特别是与道前庭和外表面的rRNA和L23r蛋白的动态相互作用.
新生链的C端区域调解着核糖体相互作用,而N端区域仍然是动态和紧的.
这些发现为共翻译蛋白折叠的机制以及特定核糖体组件在指导这一过程中的作用提供了新的见解.

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