Submitted to: Mid Atlantic Plant Molecular Biology Society Conference
Publication Type: Abstract Only
Publication Acceptance Date: November 30, 2005
Publication Date: November 30, 2005
Citation: Xu, C., Garrett, W.M., Sullivan, J., Caperna, T.J., Natarajan, S.S. 2005. Separation and identification of soybean leaf proteins by 2d-page and mass spectrometry [abstract]. Mid Atlantic Plant Molecular Biology Society Conference. Paper No. 16.
To establish a proteomic reference map for soybean leaves, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). We detected approximately 700 protein spots by 2D-PAGE using a pH 3-10 immobilized pH gradient (IPG) strip and visualization with colloidal Coomassie brilliant blue (CBB) G-250. Tryptic digests of approximately 256 protein spots were subjected to peptide mass fingerprinting by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. More than 30 protein spots were identified searching against NCBInr and SwissProt databases using the Mascot search engine. Twenty spots that were not identified by MALDI-TOF analysis were analyzed by liquid chromatography mass spectrometry (LC- MS) and were identified by searching against the NCBInr and expressed sequence tags (EST) databases. Many abundant leaf proteins are present in multiple spots. These results indicate that 2D-PAGE, combined with MALDI-TOF and LC- MS, is a sensitive and powerful technique for protein separation and identification of soybean leaf proteins.