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Title of Journal: Plant Cell Rep

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Abbravation: Plant Cell Reports

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Springer-Verlag

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DOI

10.1007/bf00915356

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1432-203X

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Proteome analysis of embryogenic cell suspensions

Authors: F C S Nogueira E F Gonçalves E S Jereissati M Santos J H Costa O B OliveiraNeto A A Soares G B Domont F A P Campos
Publish Date: 2007/02/28
Volume: 26, Issue: 8, Pages: 1333-1343
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Abstract

Using a combination of twodimensional gel electrophoresis protein mapping and mass spectrometry analysis we have established proteome reference maps of embryogenic cell suspensions of cowpea Vigna unguiculata The cell suspensions were generated from young primary leaves and contained basically proembryogenic masses which enabled us to dissect their proteome composition while eliminating the complexity of too many cell types Over 550 proteins could reproducibly be resolved over a pI range of 3–10 A total of 128 of the most abundant protein spots were excised digested ingel with trypsin and analyzed by tandem mass spectrometry This enabled the identification of 67 protein spots Two of the most abundant proteins were identified as a chitinase and as a ribonuclease belonging to the family of PR4 and PR10 proteins respectively The expression of the respective genes was confirmed by RTPCR and the pattern of deposition of the PR10 protein in cell suspensions as well as in developing cowpea seeds roots shoots and flowers were determined by Western blot experiments using synthetic antibodies raised against a 14amino acid synthetic peptide located close to the Cterminal region of the PR10 protein

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