Mason, G. G., Murray, R. Z., Pappin, D., Rivett, A. J. (July 1998) Phosphorylation of ATPase subunits of the 26S proteasome. FEBS Letters, 430 (3). pp. 269-74. ISSN 0014-5793 (Print)0014-5793 (Linking)
Abstract
The 26S proteasome complex plays a major role in the non-lysosomal degradation of intracellular proteins. Purified 26S proteasomes give a pattern of more than 40 spots on 2D-PAGE gels. The positions of subunits have been identified by mass spectrometry of tryptic peptides and by immunoblotting with subunit-specific antipeptide antibodies. Two-dimensional polyacrylamide gel electrophoresis of proteasomes immunoprecipitated from [32P]phosphate-labelled human embryo lung L-132 cells revealed the presence of at least three major phosphorylated polypeptides among the regulatory subunits as well as the C8 and C9 components of the core 20S proteasome. Comparison with the positions of the regulatory polypeptides revealed a minor phosphorylated form to be S7 (MSS1). Antibodies against S4, S6 (TBP7) and S12 (MOV34) all cross-reacted at the position of major phosphorylated polypeptides suggesting that several of the ATPase subunits may be phosphorylated. The phosphorylation of S4 was confirmed by double immunoprecipitation experiments in which 26S proteasomes were immunoprecipitated as above and dissociated and then S4 was immunoprecipitated with subunit-specific antibodies. Antibodies against the non-ATPase subunit S10, which has been suggested by others to be phosphorylated, did not coincide with the position of a phosphorylated polypeptide. Some differences were observed in the 2D-PAGE pattern of proteasomes immunoprecipitated from cultured cells compared to purified rat liver 26S proteasomes suggesting possible differences in subunit compositions of 26S proteasomes.
| Item Type: | Paper |
|---|---|
| Additional Information: | Mason, G G Murray, R Z Pappin, D Rivett, A J Wellcome Trust/United Kingdom Research Support, Non-U.S. Gov't Netherlands FEBS letters FEBS Lett. 1998 Jul 3;430(3):269-74. |
| Uncontrolled Keywords: | Adenosine Triphosphatases/ chemistry Amino Acid Sequence Animals Antibodies, Monoclonal Antibody Specificity Cell Line Cells, Cultured Electrophoresis, Gel, Two-Dimensional Humans Liver/enzymology Lung Mass Spectrometry Molecular Sequence Data Molecular Weight Peptide Hydrolases/ chemistry Phosphorylation Precipitin Tests/methods Proteasome Endopeptidase Complex Rats |
| Subjects: | bioinformatics > genomics and proteomics > design > amino acid design bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > antibodies bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > ATPase organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression > phosphorylation |
| CSHL Authors: | |
| Communities: | CSHL labs > Pappin lab |
| Depositing User: | Kathleen Darby |
| Date: | 3 July 1998 |
| Date Deposited: | 01 May 2014 13:58 |
| Last Modified: | 01 May 2014 13:58 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29917 |
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