Abstract
Proteomics is an essential tool to uncover the regulatory processes of fruit biology. In fruits with high proteolytic activity, the inhibition of endogenous proteases is key for successful protein extraction. In this chapter, we describe an efficient protocol for total protein extraction to deal with this inconvenience using pineapple pulp as an example. We corroborated the efficacy of our protein extraction protocols by carrying out nano LC-MS/MS analyses using a highly sensitive hybrid mass spectrometer. In doing so, we were able to identify over 3000 proteins in pineapple pulp. Our contribution paves the way for massive comparative proteomics scrutiny in pineapple fruits, as well as others plant tissues with high protease activity such as papaya, fig, and kiwi fruits.
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References
Li T, Wang YH, Liu JX et al (2019) Advances in genomic, transcriptomic, proteomic, and metabolomic approaches to study biotic stress in fruit crops. Crit Rev Biotechnol 39(5):680–692
Liu H, Hu M, Wang Q et al (2018) Role of papain-like cysteine proteases in plant development. Front Plant Sci 9:1717
Buono RA, Hudecek R, Nowack MK (2019) Plant proteases during developmental programmed cell death. J Exp Bot 70(7):2097–2112
Salguero-Linares J, Coll NS (2019) Plant proteases in the control of the hypersensitive response. J Exp Bot 70(7):2087–2095
Schuhmann H, Adamska I (2012) Deg proteases and their role in protein quality control and processing in different subcellular compartments of the plant cell. Physiol Plant 145:224–234
Plaxton WC (2019) Avoiding proteolysis during the extraction and purification of active plant enzymes. Plant Cell Physiol 60(4):715–724
Hellinger R, Gruber CW (2019) Peptide-based protease inhibitors from plants. Drug Discov Today 24(9):1877–1889
Niu L, Yuan H, Gong F et al (2018) Protein extraction methods shape much of the extracted proteomes. Front Plant Sci 9:802
Matagne A, Bolle L, El Mahyaoui R et al (2017) The proteolytic system of pineapple stems revisited: purification and characterization of multiple catalytically active forms. Phytochemistry 138:29–51
Acknowledgments
Esaú Bojórquez-Velázquez thanks CONACYT for the postdoctoral fellowship CVU 559120.
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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Bojórquez-Velázquez, E., Elizalde-Contreras, J.M., Zamora-Briseño, J.A., Ruiz-May, E. (2022). Efficient Protein Extraction Protocols for NanoLC-MS/MS Proteomics Analysis of Plant Tissues with High Proteolytic Activity: A Case Study with Pineapple Pulp. In: Pereira-Santana, A., Gamboa-Tuz, S.D., Rodríguez-Zapata, L.C. (eds) Plant Comparative Genomics. Methods in Molecular Biology, vol 2512. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2429-6_17
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DOI: https://doi.org/10.1007/978-1-0716-2429-6_17
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2428-9
Online ISBN: 978-1-0716-2429-6
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