Photoaffinity labeling and mass spectrometry identify ribosomal protein S3 as a potential target for hybrid polar cytodifferentiation agents Journal Article
| Authors: | Webb, Y.; Zhou, X.; Ngo, L.; Cornish, V.; Stahl, J.; Erdjument-Bromage, H.; Tempst, P.; Rifkind, R. A.; Marks, P. A.; Breslow, R.; Richon, V. M. |
| Article Title: | Photoaffinity labeling and mass spectrometry identify ribosomal protein S3 as a potential target for hybrid polar cytodifferentiation agents |
| Abstract: | The ability of a novel class of hybrid polar compounds (HPCs) to induce differentiation and consequent cessation of proliferation of transformed cells has led to their development as potential chemotherapeutic agents in the treatment of cancer. Suberoylanilide hydroxamic acid (SAHA) is a prototype of a family of hydroxamic acid based compounds (SAHA-like HPCs) that can, at micromolar concentrations, induce a variety of transformed cell lines to differentiate. The mechanism of action of the HPCs is not entirely understood. Searching for a cellular target of the SAHA-like HPCs, we synthesized a photoaffinity labeling reagent structurally based on SAHA, and probed for SAHA-binding proteins in murine erythroleukemia (MEL) cells. Photoaffinity labeling in cell free extracts identified a 32-kDa protein (p32) that was specifically labeled by the photoaffinity reagent. Cell fractionation assays localized p32 to the P100 fraction, p32 was partially purified and identified by mass spectrometry as the 40 S ribosomal protein S3. Expression of epitope-tagged S3 in bacterial lysates followed by photoaffinity labeling confirmed its specific labeling. Identification of a cytodifferentiation agent target may shed light on the mechanism by which the SAHA-like HPCs exert their antitumor effects. |
| Keywords: | human cell; cell proliferation; mass spectrometry; protein analysis; cytology; animals; mice; cell division; protein targeting; cell differentiation; antineoplastic activity; tumor cells, cultured; bacteria (microorganisms); cancer therapy; amino acid sequence; hybrid protein; protein purification; enzyme inhibitors; murinae; hydroxamic acids; cell fractionation; centrifugation; models, chemical; synthesis; ribosomal proteins; ribosome protein; imidazole derivative; cell labeling; photoaffinity labeling; amide; photochemistry; azides; affinity labels; human; priority journal; article |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 274 |
| Issue: | 20 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1999-05-14 |
| Start Page: | 14280 |
| End Page: | 14287 |
| Language: | English |
| DOI: | 10.1074/jbc.274.20.14280 |
| PUBMED: | 10318849 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 16 August 2016 -- Source: Scopus |
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