Abstract
Subcellular proteome analysis is one of the most effective ways to reduce the complexity of total proteome. With the advancement in protein extraction methodologies, it is now possible to fractionate and isolate the proteins from subcellular compartments without significant contamination from the cytoplasm and other organelles. Of the different subcellular proteomes, plasma membrane remained largely uncharacterized because of the difficulties in isolation of contamination free plasma membrane proteins. Moreover, proteome analysis in the past two decades majorly relied on the two-dimensional gel electrophoresis which showed limited protein loading ability and poor separation of highly hydrophobic plasma membrane proteins. Development of shotgun proteomics methods has facilitated the identification and quantification of hydrophobic proteins isolated from plasma membrane or other cellular membranes. Here, we present a simplified procedure for the isolation of plasma membrane proteins by a two-phase partitioning method and their identification by shotgun proteomics approach using rice as a model plant.
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References
Li B, Takahashi D, Kawamura Y et al (2018) Plasma membrane proteomics of Arabidopsis suspension-cultured cells associated with growth phase using nano-LC-MS/MS. In: Plant membrane proteomics. Springer, Cham, pp 185–194
Santoni V, Molloy M, Rabilloud T (2000) Membrane proteins and proteomics: un amour impossible? Electrophoresis 21:1054–1070
Cordwell SJ, Thingholm TE (2010) Technologies for plasma membrane proteomics. Proteomics 10:611–627
Voothuluru P, Anderson JC, Sharp RE et al (2016) Plasma membrane proteomics in the maize primary root growth zone: novel insights into root growth adaptation to water stress. Plant Cell Environ 39:2043–2054
Gupta R, Lee SE, Agrawal GK et al (2015) Understanding the plant-pathogen interactions in the context of proteomics-generated apoplastic proteins inventory. Front Plant Sci 6:352
Wang Y, Gupta R, Song W et al (2017) Label-free quantitative secretome analysis of Xanthomonas oryzae pv. Oryzae highlights the involvement of a novel cysteine protease in its pathogenicity. J Proteome 169:202–214
Wang Y, Wu J, Kim SG et al (2016) Magnaporthe oryzae-secreted protein MSP1 induces cell death and elicits defense responses in rice. Mol Plant-Microbe Interact 29:299–312
Santoni V (2007) Plant plasma membrane protein extraction and solubilization for proteomic analysis. In: Plant proteomics. Springer, Cham, pp 93–109
Meng Q, Gupta R, Min CW et al (2019) A proteomic insight into the MSP1 and flg22 induced signaling in Oryza sativa leaves. J Proteome 196:120–130
Acknowledgments
This work was supported by the National Research Foundation of Korea through Basic Research Lab (BRL) program (2018R1A4A1025158) and (2019R1A2C2085868) provided to S.T.K.
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Gupta, R., Kim, YJ., Kim, S.T. (2020). A Protocol for the Plasma Membrane Proteome Analysis of Rice Leaves. In: Jorrin-Novo, J., Valledor, L., Castillejo, M., Rey, MD. (eds) Plant Proteomics. Methods in Molecular Biology, vol 2139. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0528-8_8
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DOI: https://doi.org/10.1007/978-1-0716-0528-8_8
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