Ribosome-binding factor A (RbfA) from Staphylococcus aureus is a cold adaptation protein that is required for the growth of pathogenic cells at low temperatures (10-15°C). RbfA is involved in the processing of 16S rRNA, as well as in the assembly and stabilization of the small 30S ribosomal subunit. Structural studies of the 30S–RbfA complex will help to better understand their interaction, the mechanism of such complexes, and the fundamental process such as 30S subunit assembly that determines and controls the overall level of protein biosynthesis. This article describes protocols for preparation of RbfA and the small 30S ribosomal subunits and reconstitution and optimization of the 30S–RbfA complex to obtain samples suitable for cryo-electron microscopy studies.

中文翻译：

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用于结构研究的金黄色葡萄球菌 30S 核糖体亚基和 RbfA 因子复合物的体外重组

来自金黄色葡萄球菌的核糖体结合因子 A (RbfA) 是一种冷适应蛋白，是致病细胞在低温 (10-15°C) 下生长所必需的。RbfA 参与 16S rRNA 的加工，以及 30S 核糖体小亚基的组装和稳定。30S-RbfA 复合物的结构研究将有助于更好地了解它们的相互作用、这种复合物的机制以及基本过程，例如决定和控制蛋白质生物合成整体水平的 30S 亚基组装。本文介绍了 RbfA 和 30S 小核糖体亚基的制备方案，以及 30S-RbfA 复合物的重组和优化，以获得适用于冷冻电子显微镜研究的样品。