Status: Bibliographieeintrag
Standort: ---
Exemplare:
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| Online-Ressource |
Verfasst von: | Jovanovic, Bogdan [VerfasserIn]  |
| Schubert, Lisa [VerfasserIn]  |
| Poetz, Fabian [VerfasserIn]  |
| Stoecklin, Georg [VerfasserIn]  |
Titel: | Tagging of RPS9 as a tool for ribosome purification and identification of ribosome-associated proteins |
Verf.angabe: | Bogdan Jovanovic, Lisa Schubert, Fabian Poetz, and Georg Stoecklin |
Jahr: | 2021 |
Umfang: | 9 S. |
Fussnoten: | Gesehen am 02.06.2021 ; First published: December 30, 2020 |
Titel Quelle: | Enthalten in: Archives of biological sciences |
Ort Quelle: | Belgrade : Srpsko Biološko Društvo, 2002 |
Jahr Quelle: | 2021 |
Band/Heft Quelle: | 73(2021), 1, Seite 47-55 |
ISSN Quelle: | 1821-4339 |
Abstract: | Ribosomes, the catalytic machinery required for protein synthesis, are comprised of 4 ribosomal RNAs and about 80 ribosomal proteins in mammals. Ribosomes further interact with numerous associated factors that regulate their biogenesis and function. As mutations of ribosomal proteins and ribosome-associated proteins cause many diseases, it is important to develop tools by which ribosomes can be purified efficiently and with high specificity. Here, we designed a method to purify ribosomes from human cell lines by C-terminally tagging human RPS9, a protein of the small ribosomal subunit. The tag consists of a flag peptide and a streptavidin-binding peptide (SBP) separated by the tobacco etch virus (TEV) protease cleavage site. We demonstrate that RPS9-Flag-TEV-SBP (FTS) is efficiently incorporated into the ribosome without interfering with regular protein synthesis. Using HeLa-GFP-G3BP1 cells stably expressing RPS9-FTS or, as a negative control, mCherry-FTS, we show that complete ribosomes as well as numerous ribosome-associated proteins are efficiently and specifically purified following pull-down of RPS9-FTS using streptavidin beads. This tool will be helpful for the characterization of human ribosome heterogeneity, post-translational modifications of ribosomal proteins, and changes in ribosome-associated factors after exposing human cells to different stimuli and conditions. |
DOI: | doi:10.2298/ABS20120557J |
URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.2298/ABS20120557J |
| Volltext: http://www.doiserbia.nb.rs/Article.aspx?ID=0354-46642000057J |
| DOI: https://doi.org/10.2298/ABS20120557J |
Datenträger: | Online-Ressource |
Sprache: | eng |
Sach-SW: | affinity purification |
| evolution |
| panther |
| reveals |
| ribosome purification |
| ribosome-associated proteins |
| rps9 |
| streptavidin-binding peptide |
| stress granule |
K10plus-PPN: | 1759442445 |
Verknüpfungen: | → Zeitschrift |
Tagging of RPS9 as a tool for ribosome purification and identification of ribosome-associated proteins / Jovanovic, Bogdan [VerfasserIn]; 2021 (Online-Ressource)
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