title: Proteomic analysis of mature barley grains from C-hordein antisense lines.
abstract: Hordeins are the major storage proteins in barley grains and are responsible for their low nutritional quality. Previously, antisense C-hordein barley lines were generated and were shown to contain a more balanced amino acid composition and an altered storage protein profile. In the present study, a proteomic approach that combined two-dimensional gel electrophoresis (2-DE) and mass spectrometry was used to (1) identify the changes in the protein profile of non-storage proteins (salt soluble fraction) in antisense C-hordein barley lines (L1, L2 and L3) and (2) map the differentially expressed proteins compared to the non-transgenic control line (Hordeum vulgare cv. Golden Promise). Moreover, the changes in the proteins were correlated with the more balanced amino acid composition of these lines, with special attention to the lysine content. The results showed that suppression of C-hordein expression does not exclusively affect hordein synthesis and accumulation. The more balanced amino acid composition observed in the transgenic lines L1, L2 and L3 was an indirect result of the profound alterations in the patterns of the non-storage proteins. The observed changes included up-regulated expression of the proteins involved in stress and detoxification (L1), defence (L2 and L3), and storage globulins (L1, L2 and L3). To a lesser extent, the proteins involved in grain metabolism were also changed. Thus, the increased essential amino acids content results from changes in distinct protein sources among the three antisense C-hordein lines analyzed, although the up-regulated expression of lysine-rich proteins was consistently observed in all lines.
authors.affiliation: Departamento de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP CEP 13418-900, Brazil.
authors.forename: Daiana
authors.initials: D
authors.lastname: Schmidt
authors.affiliation: Departamento de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP CEP 13418-900, Brazil.
authors.forename: Salete Aparecida
authors.initials: SA
authors.lastname: Gaziola
authors.affiliation: Departamento de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP CEP 13418-900, Brazil.
authors.forename: Luis Felipe
authors.initials: LF
authors.lastname: Boaretto
authors.affiliation: Departamento de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP CEP 13418-900, Brazil. Electronic address: raa@usp.br.
authors.forename: Ricardo Antunes
authors.initials: RA
authors.lastname: Azevedo
mesh_terms: D002523:Edible Grain / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D005983:Glutens / Q000378:metabolism*; D001467:Hordeum / Q000737:chemistry* / Q000235:genetics; D040901:Proteomics
chemical_list: D005983:Glutens
keywords: Barley; Hordeum vulgare; Lysine-rich proteins; Mass spectrometry; Non-storage proteins; Poaceae; Two-dimensional electrophoresis