Application of free-flow IEF to identify protein candidates changing under microgravity conditions

Pietsch, Jessica and Kussian, Richard and Sickmann, Albert and Bauer, Johann and Weber, Gerhard and Nissum, Mikkel and Westphal, Kriss and Egli, Marcel and Grosse, Jirka and Schoenberger, Johann and Wildgruber, Robert and Infanger, Manfred and Grimm, Daniela (2010) Application of free-flow IEF to identify protein candidates changing under microgravity conditions. PROTEOMICS, 10 (5). pp. 904-913. ISSN 1615-9853, 1615-9861

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Abstract

Using antibody-related methods, we recently found that human thyroid cells express various proteins differently depending on whether they are cultured under normal gravity (1g) or simulated microgravity (s-mu g). In this study, we performed proteome analysis in order to identify more gravity-sensitive thyroid proteins. Cells cultured under 1g or s-mu g conditions were sonicated. Proteins released into the supernatant and those remaining in the cell fragments were fractionated by free-flow IEF. The fractions obtained were further separated by SDS-gel electrophoresis. Selected gel pieces were excised and their proteins were determined by MS. A total of 235 different proteins were found. Out of 235 proteins, 37 appeared to be first identifications in human thyroid cells. Comparing SDS gel lanes of equally numbered free-flow IEF fractions revealed similar patterns with a number of identical bands if proteins of a distinct cell line had been applied, irrespective of whether the cells had been cultured under 1g or s-mu g. Most of the identical band pairs contained identical proteins. However, the concentrations of some types of proteins were different within the two pieces of gel. Proteins that concentrated differently in such pieces of gel are considered as candidates for further investigations of gravitational sensitivity.

Item Type: Article
Uncontrolled Keywords: THYROID-CARCINOMA CELLS; HUMAN ENDOTHELIAL-CELLS; PROTEOMIC ANALYSIS; SIMULATED MICROGRAVITY; MASS-SPECTROMETRY; ELECTROPHORESIS; GROWTH; CANCER; IDENTIFICATION; ESTABLISHMENT; Cell biology; Cytoskeletal proteins; Cytosolic proteins; Free-flow electrophoresis; pl; Random positioning machine
Subjects: 600 Technology > 610 Medical sciences Medicine
Divisions: Medicine > Abteilung für Nuklearmedizin
Depositing User: Dr. Gernot Deinzer
Date Deposited: 06 Aug 2020 05:22
Last Modified: 06 Aug 2020 05:22
URI: https://pred.uni-regensburg.de/id/eprint/25132

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