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Chemoenzymatic Labeling of Proteins for Imaging in Bacterial Cells

Ho, Samuel H. and Tirrell, David A. (2016) Chemoenzymatic Labeling of Proteins for Imaging in Bacterial Cells. Journal of the American Chemical Society, 138 (46). pp. 15098-15101. ISSN 0002-7863. http://resolver.caltech.edu/CaltechAUTHORS:20161116-100355067

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Abstract

Reliable methods to determine the subcellular 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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/jacs.6b07067DOIArticle
http://pubs.acs.org/doi/abs/10.1021/jacs.6b07067PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/jacs.6b07067PublisherSupporting Information
ORCID:
AuthorORCID
Ho, Samuel H.0000-0001-7647-0752
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2016 American Chemical Society. Received: July 16, 2016; Published: November 10, 2016. The authors thank B. M. Babin, S. E. Stone, L. J. Dooling, K. P. Yuet, K. S. Burke, J. D. Bagert, and C. Kulkarni for insightful discussions. S.H.H. thanks D. K. Romney, S. Brinkmann-Chen, A. Collazo, S. Hess, A. Moridan, M. Shahgholi, C. Neubauer, and N. Dalleska for advice. S.H.H. was supported in part by a National Science Foundation Graduate Research Fellowship. This work was supported by the Caltech Center for Environmental Microbial Interactions, the Jacobs Institute for Molecular Engineering for Medicine, and the Institute for Collaborative Biotechnologies through Grant W911NF-09-0001 from the U.S. Army Research Office. The authors declare no competing financial interest.
Group:Caltech Center for Environmental Microbial Interactions (CEMI)
Funders:
Funding AgencyGrant Number
NSF Graduate Research FellowshipUNSPECIFIED
Caltech Center for Environmental Microbial Interactions (CEMI)UNSPECIFIED
Jacobs Institute for Molecular Engineering for MedicineUNSPECIFIED
Army Research Office (ARO)W911NF-09-0001
Record Number:CaltechAUTHORS:20161116-100355067
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20161116-100355067
Official Citation:Chemoenzymatic Labeling of Proteins for Imaging in Bacterial Cells Samuel H. Ho and David A. Tirrell Journal of the American Chemical Society 2016 138 (46), 15098-15101 DOI: 10.1021/jacs.6b07067
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:72050
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:16 Nov 2016 22:21
Last Modified:20 Oct 2017 22:35

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