Abstract
Proteomics is a powerful technique that allows researchers a window into how an organism responds to a mutation, a specific environment, or at a distinct point during development by quantifying relative protein abundance and posttranslational modifications. Here, we describe methods for the proteomic analysis of Arabidopsis thaliana tissue. Extraction protocols are provided for isolation of soluble, plasma membrane, and tonoplast proteins. In addition, basic analysis and quality metrics for MS/MS data are discussed. The protocols outlined have the potential to unlock new avenues of research that are not possible through basic genetics or transcriptomic approaches. By combining proteomic information with known gene regulatory patterns, researchers can gain a complete picture of how molecular pathways, such as those required for gravitropism, are initiated, regulated, and terminated.
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Acknowledgements
This work has been partially funded by NSF IOS #1147087 awarded to SEW and NASA # NNX13AM48G awarded to SEW and DRL. The authors would like to thank Dr. Frans Maathuis, University of York, for his valuable input and Dr. Sophie Alvarez, Donald Danforth Plant Science Center, St. Louis, MO, for her assistance and support with the proteomics protocols, analyses and protein quality metrics. The authors also thank Sarah Hutchinson and Marilyn Hayden for their contributions.
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Basu, P., Luesse, D.R., Wyatt, S.E. (2015). Proteomic Approaches and Their Application to Plant Gravitropism. In: Blancaflor, E. (eds) Plant Gravitropism. Methods in Molecular Biology, vol 1309. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2697-8_10
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DOI: https://doi.org/10.1007/978-1-4939-2697-8_10
Publisher Name: Humana Press, New York, NY
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