Loading...
Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes.
Barlag, Britta Beutel, Oliver Janning, Dennis Czarniak, Frederik Richter, Christian P Kommnick, Carina Göser, Vera Kurre, Rainer Fabiani, Florian
Barlag, Britta
Beutel, Oliver
Janning, Dennis
Czarniak, Frederik
Richter, Christian P
Kommnick, Carina
Göser, Vera
Kurre, Rainer
Fabiani, Florian
Files
Loading...
Open Access publication
Adobe PDF, 1.69 MB
Other Titles
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.
Citation
Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes. 2016, 6:31601 Sci Rep
Publisher
Journal
PubMed ID
PubMed Central ID
Additional Links
Embedded video
Type
Article
Language
en
Description
Series/Report no.
ISSN
2045-2322