The nutritional and cancer-protective properties of the oil extracted mechanically from the ripe fruits of Olea europaea trees are attracting constantly more attention worldwide. The preparation of high-quality protein samples from plant tissues for proteomic analysis poses many challenging problems. In this study we employed a proteomic platform based on two different extraction methods, SDS and CHAPS based protocols, followed by two precipitation protocols, TCA/acetone and MeOH precipitation, in order to increase the final number of identified proteins. The use of advanced MS techniques in combination with the Swissprot and NCBI Viridiplantae databases and TAIR10 Arabidopsis database allowed us to identify 1265 proteins, of which 22 belong to O. europaea. The application of this proteomic platform for protein extraction and identification will be useful also for other proteomic studies on recalcitrant plant/fruit tissues.
Keywords: 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; CHAPS; CMC; DTT; FDR; GO; Gel-free approach; High resolution mass spectrometry; Olive pulp; PPO; Plant proteomics; Protein extraction; RP; RPK; RT; SDS; SPE; TCA; critical micelle concentration; dithiothreitol; false discovery rate; gene ontology; polyphenol oxidase; receptor like protein kinase; reversed-phase; room temperature; sodium dodecyl sulphate; solid phase extraction; trichloroacetic acid.
Copyright © 2013. Published by Elsevier B.V.