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Probing Inner Membrane Protein Topology by Proteolysis

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Bacterial Protein Secretion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1615))

Abstract

Inner membrane proteins are inserted into the membrane via α-helices. These helices do not only constitute membrane anchors but may mediate specific interactions with membrane protein partners or participate in energetic processes. The number, location, and orientation of these helices is referred to as topology. Bitopic membrane proteins that consist of a single membrane-embedded domain connecting two soluble domains are distinguished from polytopic ones that consist of multiple membrane-spanning helices connected by extramembrane domains. Defining inner membrane protein topology could be achieved by different methods. Here we describe a protease accessibility assay that makes it possible to define topology based on digestion profiles.

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Acknowledgements

Work in EC laboratory is supported by the Centre National de la Recherche Scientifique, the Aix-Marseille Université, and grants from the Agence Nationale de la Recherche (ANR-14-CE14-0006-02 and ANR-15-CE11-0019-01). MSV is a recipient of a doctoral fellowship from the French Ministère de l’Enseignement Supérieur et de la Recherche.

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Authors and Affiliations

  1. Laboratoire d’Ingénierie des Systèmes Macromoléculaires (LISM, UMR 7255), Institut de Microbiologie de la Méditerranée (IMM), Aix-Marseille Université—Centre National de la Recherche Scientifique (CNRS), 31 Chemin Joseph Aiguier, 13402, Marseille Cedex 20, France

    Maxence S. Vincent & Eric Cascales

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  1. Maxence S. Vincent
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  2. Eric Cascales
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Correspondence to Eric Cascales .

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Editors and Affiliations

  1. Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France

    Laure Journet

  2. Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France

    Eric Cascales

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Vincent, M.S., Cascales, E. (2017). Probing Inner Membrane Protein Topology by Proteolysis. In: Journet, L., Cascales, E. (eds) Bacterial Protein Secretion Systems. Methods in Molecular Biology, vol 1615. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7033-9_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7033-9_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7031-5

  • Online ISBN: 978-1-4939-7033-9

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