Multiplex staining of 2-DE gels for an initial phosphoproteome analysis of germinating seeds and early grown seedlings from a non-orthodox specie: Quercus ilex L. subsp. ballota [Desf.] Samp

Frontiers in Plant Science
M Cristina Romero-RodríguezJesús V Jorrín-Novo

Abstract

As a preliminary step in the phosphoproteome analysis of germinating seeds (0 and 24 h after seed imbibition) and early grown seedlings (216 h after seed imbibition) from a non-orthodox sp. Quercus ilex, a multiplex (SYPRO-Ruby and Pro-Q DPS) staining of high-resolution 2-DE gels was used. By using this protocol it was possible to detect changes in protein-abundance and/or phosphorylation status. This simple approach could be a good complementary alternative to the enrichment protocols used in the search for phosphoprotein candidates. While 482 spots were visualized with SYPRO-Ruby, 222 were with Pro-Q DPS. Statistically significant differences in spot intensity were observed among samples, these corresponding to 85 SYPRO-Ruby-, 20 Pro-Q-DPS-, and 35 SYPRO-Ruby and Pro-Q-DPS-stained spots. Fifty-five phosphoprotein candidates showing qualitative or quantitative differences between samples were subjected to MALDI-TOF-TOF MS analysis, with 20 of them being identified. Identified proteins belonged to five different functional categories, namely: carbohydrate and amino acid metabolism, defense, protein folding, and oxidation-reduction processes. With the exception of a putative cyclase, the other 19 proteins had at least one orthol...Continue Reading

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Citations

Jan 8, 2017·BMC Genomics·Ramamurthy Mahalingam
Jul 28, 2018·Frontiers in Plant Science·Cristina López-HidalgoJesus V Jorrín-Novo
Feb 10, 2019·International Journal of Molecular Sciences·María-Dolores ReyJesús V Jorrín-Novo
Dec 5, 2020·International Journal of Molecular Sciences·Galina SmolikovaAndrej Frolov

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Methods Mentioned

BETA
electrophoresis
PCA
protein folding

Software Mentioned

Mascot
BLASTp
PD
PDQuest
BLAST BLAST
BLAST
Blast2GO
Quest Advance
NIA

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