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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/18186

Title: A generic approach for "shotgun" analysis of the soluble proteome of plant cell suspension cultures
Authors: De Vijlder, Thomas
VALKENBORG, Dirk
Dewaele, Debbie
Remmerie, Noor
Laukens, Kris
Witters, Erwin
Issue Date: 2015
Publisher: ELSEVIER SCIENCE BV
Citation: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 974, p. 48-56
Abstract: Cell suspension cultures from different plant species act as important model systems for studying cellular processes in plant biology and are often used as "green factories" for the production of valuable secondary metabolites and recombinant proteins. While mass spectrometry based proteome analysis techniques are ideally suited to study plant cell metabolism and other fundamental cellular processes from a birds eye perspective, they remain underused in plant studies. We describe a comprehensive sample preparation and multidimensional 'shotgun' proteomics strategy that can be generically applied to plant cell suspension cultures. This strategy was optimized and tested on an Arabidopsis thaliana ecotype Landsberg erecta culture. Furthermore, the implementation of strong cation exchange chromatography as a peptide fractionation step is elaborately tested. Its utility in mass spectrometry based proteome analysis is discussed. Using the presented analytical platform, over 13,000 unique peptides arid 2640 proteins could be identified from a single plant cell suspension sample. Finally, the experimental setup is validated using Nicotiana tabacum cv. "Bright Yellow-2" (BY-2) plant cell suspension cultures, thereby demonstrating that the presented analytical platform can also be valuable tool in proteome analysis of non-genomic model systems.
Notes: [De Vijlder, Thomas; Valkenborg, Dirk; Remmerie, Noor; Witters, Erwin] Ctr Prote CFP, B-2020 Antwerp, Belgium. [De Vijlder, Thomas; Remmerie, Noor] Univ Antwerp, Dept Biol, Lab Plant Growth & Dev, B-2020 Antwerp, Belgium. [Valkenborg, Dirk; Witters, Erwin] Vlaamse Instelling Technol Onderzoek, B-2400 Mol, Belgium. [Valkenborg, Dirk] Hasselt Univ, Interuniv Inst Biostat & Stat Bioinformat, B-3590 Diepenbeek, Belgium. [Dewaele, Debbie] Univ Antwerp, Dept Chem, B-2020 Antwerp, Belgium. [Laukens, Kris] Univ Antwerp, Antwerp Univ Hosp, Biomed Informat Res Ctr Antwerp Biomina, B-2650 Edegem, Belgium. [Laukens, Kris] Univ Antwerp, Dept Math & Comp Sci, B-2020 Antwerp, Belgium. [Witters, Erwin] Univ Antwerp, Dept Biol, Lab Ecophysiol Biochem & Toxicol, B-2020 Antwerp, Belgium.
URI: http://hdl.handle.net/1942/18186
DOI: 10.1016/j.jchromb.2014.10.024
ISI #: 000346540800007
ISSN: 1570-0232
Category: A1
Type: Journal Contribution
Validation: ecoom, 2016
Appears in Collections: Research publications

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