Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 138, Issue 46, Pages 15098-15101Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacs.6b07067
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Funding
- National Science Foundation Graduate Research Fellowship
- Caltech Center for Environmental Microbial Interactions
- Jacobs Institute for Molecular Engineering for Medicine
- Institute for Collaborative Biotechnologies from U.S. Army Research Office [W911NF-09-0001]
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Reliable methods to determine the sub cellular localization of bacterial proteins are needed for the study of prokaryotic cell biology. We describe here a simple and general technique for imaging of bacterial proteins in situ by fluorescence microscopy. The method uses the eukaryotic enzyme N-myristoyltransferase to modify the N-terminus of the protein of interest with an azido fatty acid. Subsequent strain-promoted azide-alkyne cycloaddition allows conjugation of dyes and imaging of tagged proteins by confocal fluorescence microscopy. We demonstrate the method by labeling the chemotaxis proteins Tar and CheA and the cell division proteins FtsZ and FtsA in Escherichia coli. We observe distinct spatial patterns for each of these proteins in both fixed and live cells. The method should prove broadly useful for protein imaging in bacteria.
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