Abstract
Global proteome profiling is a direct representation of the protein set in an organism, organ, tissues, or an organelle. One of the main objectives of proteomic analysis is the comparison and relative quantitation of proteins under a defined set of conditions. Two-dimensional gel electrophoresis (2-DE) has gained prominence over the last 4 decades for successfully aiding differential proteomics, providing visual confirmation of changes in protein abundance, which otherwise cannot be predicted from genome analysis. Each protein spot on 2-DE gel can be analyzed by its abundance, location, or even its presence or absence. This versatile gel-based method combines and utilizes the finest principle for separation of protein complexes by virtue of their charge and mass, visual mapping coupled with successful mass spectrometric identification of individual proteins.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Agapito-Tenfen SZ, Guerra MP, Wikmark OG, Nodari RO (2013) Comparative proteomic analysis of genetically modified maize grown under different agroecosystems conditions in Brazil. Proteome Sci 11(1)
Nouri MZ, Komatsu S (2010) Comparative analysis of soybean plasma membrane proteins under osmotic stress using gel-based and LC MS/MS-based proteomics approaches. Proteomics 10:1930–1945
Kruft V, Eubel H, Jänsch L, Werhahn W, Braun HP (2001) Proteomic approach to identify novel mitochondrial proteins in Arabidopsis. Plant Physiol 127:1694–1710
Printz B, Morais RDS, Wienkoop S, Sergeant K, Lutts S, Hausman JF, Renaut J (2015) An improved protocol to study the plant cell wall proteome. Front Plant Sci 6:237
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Barnouin K (2004) Two-dimensional gel electrophoresis for analysis of protein complexes. In: Fu H (ed) Protein-protein interactions: methods and applications 261. Humana Press Inc., New Jersey
Rabilloud T, Chevallet M, Luche S, Lelong C (2010) Two-dimensional gel electrophoresis in proteomics: past, present and future. J Proteome 73:2064–2077
Khoudoli GA, Porter IM, Blow JJ, Swedlow JR (2004) Optimisation of the two-dimensional gel electrophoresis protocol using the Taguchi approach. Proteome Sci 2:1
Liu GT, Ma L, Duan W, Wang BC, Li JH, Xu HG et al (2014) Differential proteomic analysis of grapevine leaves by iTRAQ reveals responses to heat stress and subsequent recovery. BMC Plant Biol 14:110
Rabilloud T, Vaezzadeh AR, Potier N, Lelong C, Leize-Wagner E, Chevallet M (2009) Power and limitations of electrophoretic separations in proteomics strategies. Mass Spectrom Rev 28:816–843
Bunai K, Yamane K (2005) Effectiveness and limitation of two-dimensional gel electrophoresis in bacterial membrane protein proteomics and perspectives. J Chromatogr B 815:227–236
Chevalier F (2010) Standard dyes for total protein staining in gel-based proteomic analysis. Materials 3:4784–4792
Gauci VJ, Wright EP, Coorssen JR (2011) Quantitative proteomics: assessing the spectrum of in-gel protein detection methods. J Chem Biol 4:3–29
Van den Bergh G, Arckens L (2005) Recent advances in 2D electrophoresis: an array of possibilities. Expert Rev Proteomics 2:243–252
Lilley KS, Razzaq A, Dupree P (2002) Two-dimensional gel electrophoresis: recent advances in sample preparation, detection and quantitation. Curr Opin Chem Biol 6:46–50
Santa C, Anjo SI, Manadas B (2016) Protein precipitation of diluted samples in SDS-containing buffer with acetone leads to higher protein recovery and reproducibility in comparison with TCA/acetone approach. Proteomics 6:1847–1851
Subba P, Barua P, Kumar R, Datta A, Soni KK, Chakraborty S, Chakraborty N (2013) Phosphoproteomic dynamics of chickpea (Cicer arietinum L.) reveals shared and distinct components of dehydration response. J Proteome Res 12:5025–5047
Barua P, Subba P, Lande NV, Mangalaparthi KK, Prasad TK, Chakraborty S, Chakraborty N (2016) Gel-based and gel-free search for plasma membrane proteins in chickpea (Cicer arietinum L.) augments the comprehensive data sets of membrane protein repertoire. J Proteome 143:199–208
Zychlinski AV, Gruissem W (2009) Preparation and analysis of plant and plastid proteomes by 2DE. In: Sheehan D, Tyther R (eds) Two-dimensional electrophoresis Protocols. Humana Press, New Jersey, pp 205–220
Santoni V (2007) Plant plasma membrane protein extraction and solubilization for proteomic analysis. In: Thiellement H, Zivy M, Damerval C, Méchin V (eds) Plant proteomics: methods and Protocols. Humana Press, New Jersey, pp 93–109
Zabel C, Klose J (2009) Protein extraction for 2DE. In: Sheehan D, Tyther R (eds) Two-dimensional electrophoresis protocols. Humana Press, New Jersey, pp 171–196
Stastna M, Slais K (2005) Two-dimensional gel isoelectric focusing. Electrophoresis 26:3586–3591
Acknowledgment
This work was supported by grants from the Department of Biotechnology (DBT) (BT/AGR/CG-Phase II/01/2014), India and the Council of Scientific and Industrial Research (CSIR) (38-1385/14/EMR-II), India. The authors thank Mr. Jasbeer Singh for illustrations and graphical representations in the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Barua, P., Gayen, D., Lande, N.V., Chakraborty, S., Chakraborty, N. (2017). Global Proteomic Profiling and Identification of Stress-Responsive Proteins Using Two-Dimensional Gel Electrophoresis. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 1631. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7136-7_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7136-7_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7134-3
Online ISBN: 978-1-4939-7136-7
eBook Packages: Springer Protocols