A shotgun proteomics approach was used to characterize the quinoa seed proteome. To obtain Comprehensive proteomic data from quinoa seeds three different precipitation procedures were employed: MeOH/CHCl3 /double-distilled H2O, acetone either alone or with trichloroacetic acid; the isolated proteins were then in-solution digested and the resulting peptides were analyzed by nano-liquid chromatography coupled to tandem mass spectrometry. However, since quinoa is a nonmodel plant species, only a few protein sequences are included in the most widely known protein sequence databases. To improve the data reliability a UniProt subdatabase, containing only proteins of Caryophillales order, was used. A total of 352 proteins were identified and evaluated both from a qualitative and quantitative point of view. This combined approach is certainly useful to increase the final number of identifications, but no particular class of proteins was extracted and identified in spite of the different chemistries and the different precipitation protocols. However, with respect to the other two procedures, from the relative quantitative analysis, based on the number of spectral counts, the trichloroacetic acid/acetone protocol was the best procedure for sample handling and quantitative protein extraction. This study could pave the way to further high-throughput studies on Chenopodium Quinoa

Characterization of quinoa seed proteome combining different protein precipitation techniques: Improvement of knowledge of non-model plant proteomics / Capriotti, ANNA LAURA; Cavaliere, Chiara; Piovesana, Susy; Stampachiacchiere, Serena; Ventura, Salvatore; ZENEZINI CHIOZZI, Riccardo; Lagana', Aldo. - In: JOURNAL OF SEPARATION SCIENCE. - ISSN 1615-9306. - STAMPA. - 38:6(2015), pp. 1017-1025. [10.1002/jssc.201401319]

Characterization of quinoa seed proteome combining different protein precipitation techniques: Improvement of knowledge of non-model plant proteomics

CAPRIOTTI, ANNA LAURA;CAVALIERE, CHIARA
;
PIOVESANA, SUSY;STAMPACHIACCHIERE, SERENA;VENTURA, SALVATORE;ZENEZINI CHIOZZI, RICCARDO;LAGANA', Aldo
2015

Abstract

A shotgun proteomics approach was used to characterize the quinoa seed proteome. To obtain Comprehensive proteomic data from quinoa seeds three different precipitation procedures were employed: MeOH/CHCl3 /double-distilled H2O, acetone either alone or with trichloroacetic acid; the isolated proteins were then in-solution digested and the resulting peptides were analyzed by nano-liquid chromatography coupled to tandem mass spectrometry. However, since quinoa is a nonmodel plant species, only a few protein sequences are included in the most widely known protein sequence databases. To improve the data reliability a UniProt subdatabase, containing only proteins of Caryophillales order, was used. A total of 352 proteins were identified and evaluated both from a qualitative and quantitative point of view. This combined approach is certainly useful to increase the final number of identifications, but no particular class of proteins was extracted and identified in spite of the different chemistries and the different precipitation protocols. However, with respect to the other two procedures, from the relative quantitative analysis, based on the number of spectral counts, the trichloroacetic acid/acetone protocol was the best procedure for sample handling and quantitative protein extraction. This study could pave the way to further high-throughput studies on Chenopodium Quinoa
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11573/760073
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