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Systematic characterization of nuclear proteome during apoptosis: a quantitative proteomic study by differential extraction and stable isotope labeling.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Hwang, Sun-Il Mayya, Viveka Rezaul, Karim Md |
| Copyright Year | 2006 |
| Abstract | Identification and characterization of the nuclear proteome is important for detailed understanding of multiple signaling events in eukaryotic cells. Toward this goal, we extensively characterized the nuclear proteome of human T leukemia cells by sequential extraction of nuclear proteins with different physicochemical properties using three buffer conditions. This large scale proteomic study also tested the feasibility and technical challenges associated with stable isotope labeling by amino acids in cell culture (SILAC) to uncover quantitative changes during apoptosis. Analyzing proteins from three nuclear fractions extracted from naive and apoptotic cells generated 780,530 MS/MS spectra that were used for database searching using the SEQUEST algorithm. This analysis resulted in the identification and quantification of 1,174 putative nuclear proteins. A number of known nuclear proteins involved in apoptosis as well as novel proteins not known to be part of the nuclear apoptotic machinery were identified and quantified. Consistent with SILAC-based quantifications, immunofluorescence staining of nucleus, mitochondria, and some associated proteins from both organelles revealed a dynamic recruitment of mitochondria into nuclear invaginations during apoptosis. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://mcponline.highwire.org/content/5/6/1131.full.pdf |
| Alternate Webpage(s) | http://www.mcponline.org/content/5/6/1131.full.pdf |
| PubMed reference number | 16540461v1 |
| Volume Number | 5 |
| Issue Number | 6 |
| Journal | Molecular & cellular proteomics : MCP |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Amino Acids Apoptosis Buffers Cell Culture Techniques Cell Nucleus Eukaryotic Cells Immunofluorescence assay Isotope Labeling Isotopes Mitochondria Nuclear Proteins Organelles Proteome Quantitation Staining method algorithm leukemia stable isotope |
| Content Type | Text |
| Resource Type | Article |
