Oeffinger Marlene, Wei Karen E, Rogers Richard, DeGrasse Jeffrey A, Chait Brian T, Aitchison John D, Rout Michael P
Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Nat Methods. 2007 Nov;4(11):951-6. doi: 10.1038/nmeth1101. Epub 2007 Oct 7.
The study of the dynamic interactome of cellular ribonucleoprotein (RNP) particles has been hampered by severe methodological limitations. In particular, the affinity purification of intact RNP complexes from cell lysates suffers from RNA degradation, loss of interacting macromolecules and poor overall yields. Here we describe a rapid affinity-purification method for efficient isolation of the subcomplexes that dynamically organize different RNP biogenesis pathways in Saccharomyces cerevisiae. Our method overcomes many of the previous limitations to produce large RNP interactomes with almost no contamination.
细胞核糖核蛋白（RNP）颗粒动态相互作用组的研究一直受到严重的方法学限制。特别是，从细胞裂解物中亲和纯化完整的RNP复合物存在RNA降解、相互作用大分子丢失以及总体产量低的问题。在此，我们描述了一种快速亲和纯化方法，用于高效分离酿酒酵母中动态组织不同RNP生物合成途径的亚复合物。我们的方法克服了许多先前的限制，能够产生几乎没有污染的大型RNP相互作用组。
