Self-Labeling Enzyme Tags for Translocation Analyses of Salmonella Effector Proteins
Autor(en): | Göser, V. Hensel, M. |
Stichwörter: | Bacterial Proteins; Type III Secretion Systems; Cell Line, Tumor; diffraction; Effector protein translocation; fluorescence microscopy; HeLa cell line; HeLa Cells; host pathogen interaction; Host-Pathogen Interactions; human; Humans; live cell imaging; Live-cell imaging; metabolism; nonhuman; physiology; Protein Transport; Salmonella; Salmonella enterica; Single-molecule localization and tracking (TALM) microscopy; Super-resolution microscopy; tumor cell line; Type III secretion system; type III secretion system, bacterial protein, Bacterial Proteins; Type III Secretion Systems | Erscheinungsdatum: | 2021 | Herausgeber: | Humana Press Inc. | Journal: | Methods in Molecular Biology | Volumen: | 2182 | Startseite: | 67 | Seitenende: | 82 | Zusammenfassung: | 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. © 2021, Springer Science+Business Media, LLC, part of Springer Nature. |
ISSN: | 10643745 | DOI: | 10.1007/978-1-0716-0791-6_8 | Externe URL: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85090510438&doi=10.1007%2f978-1-0716-0791-6_8&partnerID=40&md5=7529439707e05f99669f0a9dcc38389d |
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