Abstract
Plasma membrane (PM) proteins are of special interest due to their function in exchanging material and information with the external environment as well as their role in cellular regulation. In quantitative proteomic studies PM proteins are underrepresented mostly because they constitute only small percent of all membrane proteins. Strong demand is placed on plasma membrane enrichment methods. For decades two-phase partitioning Dextran T500/PEG 3350 isolation protocols were applied for many different animal and plant species and also a variety of tissue types. The typical quantity of material used in the enrichment protocols is 10–30 g of fresh weight. The main difficulty of working with in vitro cultivated plants is the low amount of material, especially when roots are examined. In addition, roots are frequently characterized by low protein concentrations. Our protocol established for roots of in vitro cultivated potato plants is adjusted to amounts of fresh weight not exceeding 7.5 g and allows studying the plasma membrane proteome by LC-MS.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Tang W (2012) Quantitative analysis of plasma membrane proteome using two-dimensional difference gel electrophoresis. Methods Mol Biol 876:67–82
Barkla JB, Pantoja O (2010) Plasma membrane and abiotic stress. Plant Cell Monogr 19:457–470
Santoni V, Molloy M, Rabilloud T (2000) Membrane proteins and proteomics: un amour impossible? Electrophoresis 21:1054–1070
Bernfur K, Larsson O, Larsson C, Gustavsson N (2013) Relative abundance of integral plasma membrane proteins in Arabidopsis leaf and root tissue determined by metabolic labeling and mass spectrometry. PLoS One 8:e71206
Komatsu S (2008) Plasma membrane proteome in Arabidopsis and rice. Proteomics 8:4137–4145
Cordwell SJ, Thingholm TE (2010) Technologies for plasma membrane proteomics. Proteomics 10:611–627
Larsson C, Widell S (2000) Isolation of plant plasma membranes and production of inside-out vesicles. Methods Enzymol 11:159–166
Schindler J, Nothwang HG (2006) Aqueous polymer two-phase systems: effective tools for plasma membrane proteomics. Proteomics 6:5409–5417
Alexandersson E, Saalbach G, Larsson C, Kjellbom P (2004) Arabidopsis plasma membrane proteomics identifies components of transport, signal transduction and membrane trafficking. Plant Cell Physiol 45:1543–1556
Mitra SK, Clouse SD, Goshe MB (2009) Enrichment and preparation of plasma membrane proteins from Arabidopsis thaliana for global proteomic analysis using liquid chromatography-tandem mass spectrometry. Methods Mol Biol 564:341–355
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
Mirza SP, Halligan BD, Greene AS, Olivier M (2007) Improved method for the analysis of membrane proteins by mass spectrometry. Physiol Genomics 30:89–94
Distler U, Kuharev J, Navarro P, Levin Y, Schild H, Tenzer S (2014) Drift time-specific collision energies enable deep-coverage data-independent acquisition proteomics. Nat Methods 11:167–170
Fischer F, Poetsch A (2006) Protein cleavage strategies for an improved analysis of the membrane proteome. Proteome Sci 4:2
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Jozefowicz, A.M., Matros, A., Witzel, K., Mock, HP. (2018). Mini-Scale Isolation and Preparation of Plasma Membrane Proteins from Potato Roots for LC/MS Analysis. In: Mock, HP., Matros, A., Witzel, K. (eds) Plant Membrane Proteomics. Methods in Molecular Biology, vol 1696. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7411-5_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7411-5_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7409-2
Online ISBN: 978-1-4939-7411-5
eBook Packages: Springer Protocols