2-DE-based proteomic analysis of common bean (Phaseolus vulgaris L.) seeds.
Common bean (Phaseolus vulgaris L.) is the most important grain legume for direct human consumption. Proteomic studies in legumes have increased significantly in the last years but few studies have been performed to date in P. vulgaris. We report here a proteomic analysis of bean seeds by two-dimensional electrophoresis (2-DE). Three different protein extraction methods (TCA-acetone, phenol and the commercial clean-up kit) were used taking into account that the extractome can have a determinant impact on the level of quality of downstream protein separation and identification. To demonstrate the quality of the 2-DE analysis, a selection of 50 gel spots was used in protein identification by mass spectrometry (MALDI-TOF MS and MALDI-TOF/TOF). The results showed that a considerable proportion of spots (70%) were identified in spite of incomplete genome/protein databases for bean and other legume species. Most identified proteins corresponded to storage protein, carbohydrate metabolism, defense and stress response, including proteins highly abundant in the seed of P. vulgaris such as the phaseolin, the phytohemagglutinin and the lectin-related α-amylase inhibitor.
Department of Plant Genetic Resources, Misión Biológica de Galicia-CSIC, Pontevedra, Spain. mfuente@mbg.cesga.es
M
M
De La Fuente
A
A
Borrajo
J
J
Bermúdez
M
M
Lores
J
J
Alonso
M
M
López
M
M
Santalla
A M
AM
De Ron
C
C
Zapata
G
G
Alvarez
D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D004791:Enzyme Inhibitors / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D018745:Genome, Plant; D006801:Humans; D027805:Phaseolus / Q000737:chemistry* / Q000378:metabolism; D010835:Phytohemagglutinins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012639:Seeds / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D004791:Enzyme Inhibitors; D010835:Phytohemagglutinins; D010940:Plant Proteins; D020543:Proteome