Identification of proteins binding the native tubulin dimer
| Autor: | Gache, Vincent, Louwagie, Mathilde, Garin, Jérôme, Caudron, Nicolas, Lafanechere, Laurence, Valiron, Odile |
|---|---|
| Přispěvatelé: | Organisation Fonctionnelle du Cytosquelette, Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR27, Laboratoire de chimie des protéines, Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), La Ligue Nationale Contre le Cancer ('équipe labellisée Ligue') to DJ |
| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
MESH: Humans
MESH: Protein Structure Quaternary macromolecular substances MESH: Tubulin MESH: Chromatography Affinity MESH: HSC70 Heat-Shock Proteins MESH: Tyrosine MESH: Hela Cells MESH: Cattle MESH: Microtubule-Associated Proteins MESH: Centrifugation Density Gradient MESH: Dimerization MESH: Protein Binding MESH: Animals [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology MESH: HSP70 Heat-Shock Proteins |
| Zdroj: | Biochemical and Biophysical Research Communications Biochemical and Biophysical Research Communications, Elsevier, 2005, 327 (1), pp.35-42. ⟨10.1016/j.bbrc.2004.11.138⟩ Biochemical and Biophysical Research Communications, 2005, 327 (1), pp.35-42. ⟨10.1016/j.bbrc.2004.11.138⟩ |
| ISSN: | 0006-291X 1090-2104 |
| DOI: | 10.1016/j.bbrc.2004.11.138⟩ |
| Popis: | International audience; Microtubules play an essential role in eukaryotic cells, where they perform a wide variety of functions. In this paper, we describe the characterization of proteins associated to tubulin dimer in its native form, using affinity chromatography and mass spectrometry. We used an immunoaffinity column with coupled-monoclonal antibody directed against the alpha-tubulin C-terminus. Tubulin was first loaded onto the column, then interphase and mitotic cell lysates were chromatographed. Tubulin-binding proteins were eluted using a peptide mimicking the alpha-tubulin C-terminus. Elution fractions were analyzed by SDS-PAGE, and a total of 14 proteins were identified with high confidence by mass spectrometry. These proteins could be grouped in four classes: known tubulin-binding proteins, one microtubule-associated protein, heat shock proteins, and proteins that were not shown previously to bind tubulin dimer or microtubules. |
| Databáze: | OpenAIRE |
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