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Chemical tools for protein imaging in live bacterial cells

Ho, Samuel and Tirrell, David (2018) Chemical tools for protein imaging in live bacterial cells. In: 255th American Chemical Society National Meeting & Exposition, March 18-22, 2018, New Orleans, LA.

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Reliable methods to det. the subcellular localization of bacterial proteins are needed for the study of prokaryotic cell biol. We report a simple and general technique for imaging 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 cycloaddn. allows conjugation of dyes and imaging of tagged proteins by confocal fluorescence microscopy. This method is demonstrated through labeling of the chemotaxis proteins Tar and CheA and the cell division proteins FtsZ and FtsA in Escherichia coli. Distinct spatial patterns for each of these proteins are obsd. in both fixed and live cells. Furthermore, we describe a photoswitchable fluorescent reporter to effect super-resoln. imaging of bacterial chemotaxis proteins. The methods described herein should prove broadly useful for protein imaging in bacteria.

Item Type:Conference or Workshop Item (Paper)
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URLURL TypeDescription Website
Ho, Samuel0000-0001-7647-0752
Tirrell, David0000-0003-3175-4596
Additional Information:© 2018 American Chemical Society.
Record Number:CaltechAUTHORS:20180412-161333505
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:85801
Deposited By: Tony Diaz
Deposited On:12 Apr 2018 23:49
Last Modified:03 Oct 2019 19:35

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