Nature Methods
- 4, 951 - 956 (2007)
Published online: 7 October 2007; | doi:10.1038/nmeth1101
Comprehensive analysis of diverse ribonucleoprotein complexesMarlene Oeffinger1, Karen E Wei1, Richard Rogers2, Jeffrey A DeGrasse1, Brian T Chait1, John D Aitchison2 & Michael P Rout11
Rockefeller University, 1230 York Avenue, New York, New York 10021, USA. 2
Institute for Systems Biology, 1141 N. 34th St., Seattle, Washington 98103, USA.
Correspondence should be addressed to Michael P Rout rout@rockefeller.edu 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.
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