Pietsch, Jessica and Bauer, Johann and Weber, Gerhard and Nissum, Mikkel and Westphal, Kriss and Egli, Marcel and Grosse, Jirka and Schoenberger, Johann and Eilles, Christoph and Infanger, Manfred and Grimm, Daniela (2011) Proteome Analysis of Thyroid Cancer Cells After Long-Term Exposure to a Random Positioning Machine. MICROGRAVITY SCIENCE AND TECHNOLOGY, 23 (4). pp. 381-390. ISSN 0938-0108, 1875-0494
Full text not available from this repository. (Request a copy)Abstract
Annulling gravity during cell culturing triggers various types of cells to change their protein expression in a time dependent manner. We therefore decided to determine gravity sensitive proteins and their period of sensitivity to the effects of gravity. In this study, thyroid cancer cells of the ML-1 cell line were cultured under normal gravity (1 g) or in a random positioning machine (RPM), which simulated near weightlessness for 7 and 11 days. Cells were then sonicated and proteins released into the supernatant were separated from those that remained attached to the cell fragments. Subsequently, both types of proteins were fractionated by free-flow isoelectric focussing (FF-IEF). The fractions obtained were further separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to which comparable FF-IEF fractions derived from cells cultured either under 1 g or on the RPM had been applied side by side. The separation resulted in pairs of lanes, on which a number of identical bands were observed. Selected gel pieces were excised and their proteins determined by mass spectrometry. Equal proteins from cells cultured under normal gravity and the RPM, respectively, were detected in comparable gel pieces. However, many of these proteins had received different Mascot scores. Quantifying heat shock cognate 71 kDa protein, glutathione S-transferase P, nucleoside diphosphate kinase A and annexin-2 by Western blotting using whole cell lysates indicated usefulness of Mascot scores for selecting the most efficient antibodies.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HUMAN ENDOTHELIAL-CELLS; MASS-SPECTROMETRY DATA; SIMULATED MICROGRAVITY; GENE-EXPRESSION; CARCINOMA CELLS; GROWTH; APOPTOSIS; DIFFERENTIATION; WEIGHTLESSNESS; INHIBITOR; Free-flow electrophoresis; Isoelectric point; Random positioning machine; Microgravity; Mass spectrometry |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Abteilung für Nuklearmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 27 May 2020 06:52 |
| Last Modified: | 27 May 2020 06:52 |
| URI: | https://pred.uni-regensburg.de/id/eprint/19853 |
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