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Self-Labeling Enzyme Tags for Translocation Analyses of Salmonella Effector Proteins

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Salmonella

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2182))

Abstract

Salmonella enterica is an invasive, facultative intracellular pathogen with a highly sophisticated intracellular lifestyle. Invasion and intracellular proliferation are dependent on the translocation of effector proteins by two distinct type III secretion systems (T3SS) into the host cell. To unravel host-pathogen interactions, dedicated imaging techniques visualizing Salmonella effector proteins during the infection are essential. Here we describe a new approach utilizing self-labeling enzyme (SLE) tags as a universal labeling tool for tracing effector proteins. This method is able to resolve the temporal and spatial dynamics of effector proteins in living cells. The method is applicable to conventional confocal fluorescence microscopy, but also to tracking and localization microscopy (TALM), and super-resolution microscopy (SRM) of single molecules, allowing the visualization of effector proteins beyond the optical diffraction limit.

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Acknowledgments

This work was supported by grant HE 1964/18-2 and SFB 944 project Z to M.H. We like to thank Jacob Piehler (Div. Biophysics) and Rainer Kurre (iBiOs) for continuous support and fruitful discussions, as well as Christian P. Richter (Div. Biophysics) for providing the localization and tracking software and the support during data analysis.

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Authors and Affiliations

  1. Abteilung Mikrobiologie, Fachbereich Biologie/Chemie, Universität Osnabrück, Osnabrück, Germany

    Vera Göser

  2. Abteilung Mikrobiologie, CellNanOs - Center for Cellular Nanoanalytics Osnabrueck, Universitaet Osnabrueck, Osnabrueck, Germany

    Michael Hensel

Authors
  1. Vera Göser
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  2. Michael Hensel
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Correspondence to Michael Hensel .

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Editors and Affiliations

  1. Department of Veterinary Pathobiology, University of Missouri-Columbia, Columbia, MO, USA

    Heide Schatten

1 Electronic Supplementary Material

Data Transformation and calculation of UC50 and IC50 data (MP4 3676 kb)

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Göser, V., Hensel, M. (2021). Self-Labeling Enzyme Tags for Translocation Analyses of Salmonella Effector Proteins. In: Schatten, H. (eds) Salmonella. Methods in Molecular Biology, vol 2182. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0791-6_8

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  • DOI: https://doi.org/10.1007/978-1-0716-0791-6_8

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