Hung Victoria, Susanto Teodorus Theo, Barna Maria
Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
STAR Protoc. 2025 Sep 1;6(3):104052. doi: 10.1016/j.xpro.2025.104052.
Ribosome-associated proteins (RAPs) enable modulation of gene expression at the level of mRNA translation in response to cellular needs. Here, we describe a method called ribosome-associated protein identification by affinity to sulfhydryl-charged resin (RAPIDASH) for tag-free isolation of RAP-bound ribosomes from mammalian samples for mass spectrometry-based proteomics. Samples are first lysed and then undergo sucrose cushion ultracentrifugation and subsequent chromatography using a sulfhydryl-charged resin. While RAPIDASH is optimized for mammalian samples, we expect that it can be adapted for non-mammalian samples. For complete details on the use and execution of this protocol, please refer to Susanto et al..
核糖体相关蛋白（RAPs）能够根据细胞需求在mRNA翻译水平调节基因表达。在此，我们描述了一种称为基于巯基电荷树脂亲和力的核糖体相关蛋白鉴定法（RAPIDASH），用于从哺乳动物样本中无标签分离与RAP结合的核糖体，以进行基于质谱的蛋白质组学研究。首先对样本进行裂解，然后进行蔗糖垫层超速离心，随后使用巯基电荷树脂进行色谱分析。虽然RAPIDASH针对哺乳动物样本进行了优化，但我们预计它也可适用于非哺乳动物样本。有关此方案的使用和执行的完整详细信息，请参考苏桑托等人的研究。
