Quantitative proteome analysis of detergent-resistant membranes identifies the differential regulation of protein kinase C isoforms in apoptotic T cells
Solstad, T; Bjørgo, E; Koehler, CJ; Strozynski, M; Torgersen, KM; Taskén, K; Thiede, B
HERO ID
1055762
Reference Type
Journal Article
Year
2010
Language
English
PMID
| HERO ID | 1055762 |
|---|---|
| In Press | No |
| Year | 2010 |
| Title | Quantitative proteome analysis of detergent-resistant membranes identifies the differential regulation of protein kinase C isoforms in apoptotic T cells |
| Authors | Solstad, T; Bjørgo, E; Koehler, CJ; Strozynski, M; Torgersen, KM; Taskén, K; Thiede, B |
| Journal | Proteomics |
| Volume | 10 |
| Issue | 15 |
| Page Numbers | 2758-2768 |
| Abstract | Several lines of evidence suggest that detergent-resistant membranes (DRMs) (also known as lipid rafts and glycosphingolipid-enriched microdomains) may have a role in signaling pathways of apoptosis. Here, we developed a method that combines DRMs isolation and methanol/chloroform extraction with stable isotope labeling with amino acids in cell culture-based quantitative proteome analysis of DRMs from control and cisplatin-induced apoptotic Jurkat T cells. This approach enabled us to enrich proteins with a pivotal role in cell signaling of which several were found with increased or decreased amounts in DRMs upon induction of apoptosis. Specifically, we show that three isoforms of protein kinase C (PKC) are regulated differently upon apoptosis. Although PKC alpha which belongs to the group of conventional PKCs is highly up-regulated in DRMs, the levels of two novel PKCs, PKC eta and PKC theta, are significantly reduced. These alterations/differences in PKC regulation are verified by immunoblotting and confocal microscopy. In addition, a specific enrichment of PKC alpha in apoptotic blebs and buds is shown. Furthermore, we observe an increased expression of ecto-PKC alpha as a result of exposure to cisplatin using flow cytometry. Our results demonstrate that in-depth proteomic analysis of DRMs provides a tool to study differential localization and regulation of signaling molecules important in health and disease. |
| Doi | 10.1002/pmic.201000164 |
| Pmid | 20486122 |
| Wosid | WOS:000281054300003 |
| Is Certified Translation | No |
| Dupe Override | No |
| Comments | Source: Web of Science 000281054300003 |
| Is Public | Yes |
| Language Text | English |
| Keyword | Apoptosis; Cell biology; Detergent-resistant membranes; Lipid rafts; Protein kinase C; Quantitative proteomics |
| Is Qa | No |