Analytical and Bioanalytical Chemistry

, Volume 402, Issue 1, pp 299–314

New insights on proteomics of transgenic soybean seeds: evaluation of differential expressions of enzymes and proteins

  • Herbert S. Barbosa
  • Sandra C. C. Arruda
  • Ricardo A. Azevedo
  • Marco A. Z. Arruda
Original Paper

DOI: 10.1007/s00216-011-5409-1

Cite this article as:
Barbosa, H.S., Arruda, S.C.C., Azevedo, R.A. et al. Anal Bioanal Chem (2012) 402: 299. doi:10.1007/s00216-011-5409-1

Abstract

This work reports the evaluation of differentially expressed enzymes and proteins from transgenic and nontransgenic soybean seeds. Analysis of malondialdehyde, ascorbate peroxidase (EC 1.11.1.11), glutathione reductase (EC 1.6.4.2), and catalase (EC 1.11.1.6) revealed higher levels (29.8, 30.6, 71.4, and 35.3%, respectively) in transgenic seeds than in nontransgenic seeds. Separation of soybean seed proteins was done by two-dimensional polyacrylamide gel electrophoresis, and 192 proteins were identified by matrix-assisted laser desorption/ionization (MALDI) quadrupole time-of-flight (QTOF) mass spectrometry (MS) and electrospray ionization (ESI) QTOF MS. Additionally, the enzyme CP4 EPSPS, involved in the genetic modification, was identified by enzymatic digestions using either trypsin or chymotrypsin and ESI-QTOF MS/MS for identification. From the proteins identified, actin fragment, cytosolic glutamine synthetase, glycinin subunit G1, and glycine-rich RNA-binding protein were shown to be differentially expressed after analysis using the two-dimensional difference gel electrophoresis technique, and applying a regulator factor of 1.5 or greater.

Figure

Application of in gel electrophoretic techniques for evaluation of transgenic soybean seeds from differential expressions of enzymes and proteins

Keywords

Two-dimensional difference gel electrophoresis Mass spectrometry Reactive oxygen species Genetically modified organisms 

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Herbert S. Barbosa
    • 1
    • 2
  • Sandra C. C. Arruda
    • 3
  • Ricardo A. Azevedo
    • 3
  • Marco A. Z. Arruda
    • 1
    • 2
  1. 1.Spectrometry, Sample Preparation and Mechanization Group - GEPAM, Institute of ChemistryUniversity of Campinas - UNICAMPCampinasBrazil
  2. 2.National Institute of Science and Technology for Bioanalytics, Institute of ChemistryUniversity of Campinas – UNICAMPCampinasBrazil
  3. 3.Laboratory of Biochemistry and Genetics of Plants, Department of GeneticsEscola Superior de Agricultura Luiz de Queiroz, ESALQ, USPPiracicabaBrazil