Techniques in Plant Proteomics
Plant proteomics is a relatively new research field that focuses on large-scale functional analysis of plant proteins. This new research field has already demonstrated immense potential for getting significantly deeper insight into the functional interaction of plant proteins and their roles in plant growth and development. The knowledge of key proteins responsible for valuable crop traits in the context of expression of other involved proteins plays pivotal role in discovery phase of crop improvement effort. This chapter introduces the field of plant proteomics in practical manner. Set of six figures helps to understand principles of protein extraction, separation and identification by mass spectrometry. Protein and peptide separation is introduced in greater details taking into account “gel-based” and “gel-free” methods.
KeywordsSodium Dodecyl Sulphate Protein Identification Peptide Mass Fingerprinting Poppy Seed Column Packing Material
The work was supported by FP7 of the European Union (MIRG-CT-2007-200165). This chapter reflects only the author’s views and the Community is not liable for any use that might be made of information contained herein.
- Adams P, Fowler R, Howell G, Kinsella N, Skipp P, Coote P, O’Connor CD (1999) Defining protease specificity with proteomics: a protease with a dibasic amino acid recognition motif is regulated by a two-component signal transduction system in Salmonella. Electrophoresis 20:2241–2247CrossRefPubMedGoogle Scholar
- Alexandrov ML,Gall LN, Krasnov NV, Nikolaev VI, Pavlenko VA, Shkurov VA (1984) Ion extraction from solutions at atmospheric pressure – a method for mass-spectrometric analysis for mass-spectrometric analysis of bioorganic substances. Dokl Akad Nauk SSSR 277:379–383Google Scholar
- Corbett JM, Dunn MJ, Posch A et al. (1994) Positional reproducibility of protein spots in two-dimensional polyacrylamide gel electrophoresis using immobilised pH gradient isoelectric focusing in the first dimension, An interlaboratory comparison. Electrophoresis 15:1205–1211CrossRefPubMedGoogle Scholar
- Davidssona P, Sjogrena M (2005) The use of proteomics in biomarker discovery in neurodegenerative diseases. Dis Markers 21:81–92Google Scholar
- Rehm H (2006) Protein biochemistry and proteomics. Academic, LondonGoogle Scholar
- Ross PL, Huang YN, Marchese JN, Williamson B, Parker K, Hattan S, Khainovski N, Pillai S, Dey S, Daniels S, Purkayastha S, Juhasz P, Martin S, Bartlet-Jones M, He F, Jacobson A, Pappin DJ (2004) Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. Mol Cell Proteomics 12:1154–1169Google Scholar
- Tishchenko G, Dybal J, Mészárosová K et al. (2002) Purification of the specific immunoglobulin G1 by immobilized metal ion affinity chromatography using nickel complexes of chelating porous and nonporous polymeric sorbents based on poly(methacrylic esters) effect of polymer structure. J Chromatogr A 954:115–126CrossRefPubMedGoogle Scholar
- Wehr T (2001) Separation Technology in Proteomics. LCGC 19:702–711Google Scholar