Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes.

2.50
Hdl Handle:
http://hdl.handle.net/10033/620079
Title:
Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes.
Authors:
Barlag, Britta; Beutel, Oliver; Janning, Dennis; Czarniak, Frederik; Richter, Christian P; Kommnick, Carina; Göser, Vera; Kurre, Rainer; Fabiani, Florian; Erhardt, Marc; Piehler, Jacob; Hensel, Michael
Abstract:
The investigation of the subcellular localization, dynamics and interaction of proteins and protein complexes in prokaryotes is complicated by the small size of the cells. Super-resolution microscopy (SRM) comprise various new techniques that allow light microscopy with a resolution that can be up to ten-fold higher than conventional light microscopy. Application of SRM techniques to living prokaryotes demands the introduction of suitable fluorescent probes, usually by fusion of proteins of interest to fluorescent proteins with properties compatible to SRM. Here we describe an approach that is based on the genetically encoded self-labelling enzymes HaloTag and SNAP-tag. Proteins of interest are fused to HaloTag or SNAP-tag and cell permeable substrates can be labelled with various SRM-compatible fluorochromes. Fusions of the enzyme tags to subunits of a type I secretion system (T1SS), a T3SS, the flagellar rotor and a transcription factor were generated and analysed in living Salmonella enterica. The new approach is versatile in tagging proteins of interest in bacterial cells and allows to determine the number, relative subcellular localization and dynamics of protein complexes in living cells.
Affiliation:
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes. 2016, 6:31601 Sci Rep
Journal:
Scientific reports
Issue Date:
2016
URI:
http://hdl.handle.net/10033/620079
DOI:
10.1038/srep31601
PubMed ID:
27534893
Type:
Article
Language:
en
ISSN:
2045-2322
Appears in Collections:
publications of the junior research group infection biology of Salmonella (IBIS)

Full metadata record

DC FieldValue Language
dc.contributor.authorBarlag, Brittaen
dc.contributor.authorBeutel, Oliveren
dc.contributor.authorJanning, Dennisen
dc.contributor.authorCzarniak, Frederiken
dc.contributor.authorRichter, Christian Pen
dc.contributor.authorKommnick, Carinaen
dc.contributor.authorGöser, Veraen
dc.contributor.authorKurre, Raineren
dc.contributor.authorFabiani, Florianen
dc.contributor.authorErhardt, Marcen
dc.contributor.authorPiehler, Jacoben
dc.contributor.authorHensel, Michaelen
dc.date.accessioned2016-09-13T09:40:47Z-
dc.date.available2016-09-13T09:40:47Z-
dc.date.issued2016-
dc.identifier.citationSingle molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes. 2016, 6:31601 Sci Repen
dc.identifier.issn2045-2322-
dc.identifier.pmid27534893-
dc.identifier.doi10.1038/srep31601-
dc.identifier.urihttp://hdl.handle.net/10033/620079-
dc.description.abstractThe investigation of the subcellular localization, dynamics and interaction of proteins and protein complexes in prokaryotes is complicated by the small size of the cells. Super-resolution microscopy (SRM) comprise various new techniques that allow light microscopy with a resolution that can be up to ten-fold higher than conventional light microscopy. Application of SRM techniques to living prokaryotes demands the introduction of suitable fluorescent probes, usually by fusion of proteins of interest to fluorescent proteins with properties compatible to SRM. Here we describe an approach that is based on the genetically encoded self-labelling enzymes HaloTag and SNAP-tag. Proteins of interest are fused to HaloTag or SNAP-tag and cell permeable substrates can be labelled with various SRM-compatible fluorochromes. Fusions of the enzyme tags to subunits of a type I secretion system (T1SS), a T3SS, the flagellar rotor and a transcription factor were generated and analysed in living Salmonella enterica. The new approach is versatile in tagging proteins of interest in bacterial cells and allows to determine the number, relative subcellular localization and dynamics of protein complexes in living cells.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleSingle molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalScientific reportsen

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