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N-Myristoyl Transferase (NMT)-Catalyzed Labeling of Bacterial Proteins for Imaging in Fixed and Live Cells

Ho, Samuel H. and Tirrell, David A. (2019) N-Myristoyl Transferase (NMT)-Catalyzed Labeling of Bacterial Proteins for Imaging in Fixed and Live Cells. In: Enzyme-Mediated Ligation Methods. Methods in Molecular Biology. No.2012. Humana Press , New York, NY, pp. 315-326. ISBN 978-1-4939-9545-5. http://resolver.caltech.edu/CaltechAUTHORS:20190610-084517342

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Abstract

Methods for selective protein imaging are critical for elucidating how cells orchestrate fundamental biological processes. We recently developed a chemoenzymatic method to modify bacterial proteins in situ for fluorescence imaging using N-myristoyl transferase (NMT). Target proteins outfitted with an N-terminal NMT recognition sequence are covalently modified with an azido fatty acid. Subsequent strain-promoted azide–alkyne cycloaddition allows for conjugation to cell-permeant fluorophores and imaging by fluorescence microscopy. Here we describe sample preparation and labeling protocols for imaging bacterial proteins in fixed and live cells.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
https://doi.org/10.1007/978-1-4939-9546-2_16DOIArticle
ORCID:
AuthorORCID
Ho, Samuel H.0000-0001-7647-0752
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2019 Springer Science+Business Media, LLC, part of Springer Nature. First Online 04 June 2019.
Subject Keywords:N-Myristoyl transferase (NMT); 12-Azidododecanoic acid; Bicyclononyne–BODIPY; Escherichia coli; Site-specific; Protein labeling; Fluorescence imaging
Record Number:CaltechAUTHORS:20190610-084517342
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20190610-084517342
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:96228
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:10 Jun 2019 16:12
Last Modified:10 Jun 2019 16:12

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