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Proteomics in globe artichoke: Protein extraction and sample complexity reduction by PEG fractionation
Author(s) -
Acquadro Alberto,
Falvo Sara,
Mila Silvia,
Giuliano Albo Alessandra,
Comino Cinzia,
Moglia Andrea,
Lanteri Sergio
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200800549
Subject(s) - rubisco , fractionation , proteomics , proteome , cell fractionation , chromatography , biochemistry , chemistry , oxygenase , pyruvate carboxylase , biology , botany , photosynthesis , enzyme , gene
Here, we report the first leaf proteome analysis for globe artichoke. Three protein extraction protocols were tested and a reproducible Mg/NP‐40‐based method was established. Ribulose‐1,5‐biphosphate carboxylase‐oxygenase (RuBisCO) is a highly abundant leaf protein, and its presence masks co‐localizing, less abundant proteins. To remove RuBisCO from the sample, and thereby improve spot resolution, a PEG fractionation approach was elaborated. 2‐DE profiles of various PEG fractions showed that the fractionation procedure was successful in excluding most of the RuBisCO, allowing for the detection of many low‐abundance proteins. Western blot analysis was able to confirm the reduction in RuBisCO content achieved by PEG fractionation. In all, 841 distinct protein spots were detected, and 40 of these, selected from the RuBisCO region of the 2‐DE profile, were successfully identified by MS. A number of homologues of these proteins also co‐localize with RuBisCO in Arabidopsis thaliana.