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Comprehensive analysis of diverse ribonucleoprotein complexes.
- Source :
-
Nature methods [Nat Methods] 2007 Nov; Vol. 4 (11), pp. 951-6. Date of Electronic Publication: 2007 Oct 07. - Publication Year :
- 2007
-
Abstract
- 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.
- Subjects :
- DNA-Binding Proteins analysis
DNA-Binding Proteins genetics
Electrophoresis, Polyacrylamide Gel
Green Fluorescent Proteins analysis
Green Fluorescent Proteins genetics
Immunosorbent Techniques
Mass Spectrometry
Nuclear Proteins analysis
Nuclear Proteins genetics
Nucleocytoplasmic Transport Proteins analysis
Nucleocytoplasmic Transport Proteins genetics
Oligonucleotide Array Sequence Analysis
Porins
RNA Cap-Binding Proteins
RNA, Fungal analysis
RNA, Messenger analysis
RNA, Ribosomal analysis
RNA-Binding Proteins analysis
RNA-Binding Proteins genetics
Ribonucleoproteins genetics
Ribosomal Proteins analysis
Ribosomal Proteins genetics
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins genetics
Staphylococcal Protein A analysis
Staphylococcal Protein A genetics
Ribonucleoproteins analysis
Saccharomyces cerevisiae metabolism
Saccharomyces cerevisiae Proteins analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1548-7091
- Volume :
- 4
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Nature methods
- Publication Type :
- Academic Journal
- Accession number :
- 17922018
- Full Text :
- https://doi.org/10.1038/nmeth1101