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Identification of selenium in the leaf protein of sunflowers by a combination of 2D-PAGE and laser ablation ICP-MS |
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| Authors: | Marcelo Anselmo Oseas da Silva Marco Aurélio Zezzi Arruda |
| Institution: | 1. Spectrometry, Sample Preparation and Mechanization Group (GEPAM) and National Institute of Science and Technology for Bioanalytics – Institute of Chemistry, University of Campinas – Unicamp, PO Box 6154, 13083-970, Campinas, SP, Brazil |
| Abstract: | We report on a method for the identification of selenium-containing proteins in an extract of sunflower leafs. It is based on the separation of the proteins by 2-dimensional gel electrophoresis, followed by detection of selenium via laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The laser system was operated in a raster mode at 100?μm?s-1 and proved to be an efficient alternative in the search for selenoproteins in the spots of the gels. The instrumental parameters were optimized in terms of plasma energy and application of optimal reaction cell conditions, and the detection of the mass 80Se16O+ which enabled the elimination of interfering species. Selenium was identified in 9.6% of the analyzed spots, indicating its random incorporation into the primary structure of the proteins. This work describes the detection of selenium in sunflower leaf proteins from plants irrigated with selenite ions by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) after protein extraction and separation through two-dimensional gel electrophoresis. |
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