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Identification of Effectors: Precipitation of Supernatant Material

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Bacterial Secretion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2715))

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Abstract

Bacterial secretion systems allow the transport of proteins, called effectors, as well as external machine components in the extracellular medium or directly into target cells. Comparison of the secretome, i.e., the proteins released in the culture medium, of wild-type and mutant cells provides information on the secretion profile. In addition, mass spectrometry analyses of the culture supernatant of bacteria grown in liquid culture under secreting conditions allow the identification of secretion systems substrates. Upon identification of the substrates, the secretion profile serves as a tool to test the functionality of secretion systems. Here, we present a classical method used to concentrate the culture supernatant, based on TCA precipitation.

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Acknowledgments

Work of L.J. was supported by the Centre National de la Recherche Scientifique, the Aix-Marseille Université, and grants from the Agence Nationale de la Recherche (ANR-14-CE14-0006 and ANR-18-CE15-0013). PhD studies of N.F. were supported by the ANR-14-CE14-0006 grant.

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Correspondence to Laure Journet .

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Flaugnatti, N., Journet, L. (2024). Identification of Effectors: Precipitation of Supernatant Material. In: Journet, L., Cascales, E. (eds) Bacterial Secretion Systems . Methods in Molecular Biology, vol 2715. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3445-5_32

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  • DOI: https://doi.org/10.1007/978-1-0716-3445-5_32

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3444-8

  • Online ISBN: 978-1-0716-3445-5

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