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  Substrates for improved live-cell fluorescence labeling of SNAP-tag.

Correa, I. R., Baker, B., Zhang, A., Sun, L., Provost, C. R., Lukinavicius, G., et al. (2013). Substrates for improved live-cell fluorescence labeling of SNAP-tag. Current Pharmaceutical Design, 19(30), 5414-5420. doi:10.2174/1381612811319300011.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0001-DE8B-9 Version Permalink: http://hdl.handle.net/21.11116/0000-0001-DE8E-6
Genre: Journal Article

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 Creators:
Correa, I. R., Author
Baker, B., Author
Zhang, A., Author
Sun, L., Author
Provost, C. R., Author
Lukinavicius, G.1, Author              
Reymond, L., Author
Johnsson, K., Author
Xu, M. Q., Author
Affiliations:
1Laboratory of Chromatin Labeling and Imaging, Max Planck Institute for Biophysical Chemistry, Max Planck Society, ou_2616691              

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Free keywords: Cell imaging, covalent labeling, fluorescent probes, protein modifications
 Abstract: The SNAP-tag labeling technology provides a simple, robust, and versatile approach to the imaging of fusion proteins for a wide range of experimental applications. Owing to the specific and covalent nature of the labeling reaction, SNAP-tag is well suited for the analysis and quantification of fused target protein using fluorescence microscopy techniques. In this report, we present our most recent findings on the labeling of SNAP-tag fusion proteins both in vitro and in cell culture with SNAP-tag substrates derived from single regioisomers of carboxyrhodamine dyes. Carboxyrhodamines are invaluable fluorescent dyes for biotechnology applications including DNA sequencing, detection on microarrays, and fluorescence in situ hybridization. We found that SNAP-tag reacts preferentially with the 6-positional regioisomer of carboxyrhodamine fluorescent dyes, whereas the 5-regioisomer predominantly contributes to background fluorescence. Our experimental study also indicates that benzylchloropyrimidine (CP) conjugates of 6-carboxyrhodamines exhibit a dramatic increase in the signal-to-noise ratio of fluorescently labeled cellular proteins compared to the benzylguanine (BG) conjugates, presumably due to higher cell permeability. These new SNAP-tag substrates based on pure 6-regioisomers can significantly improve fluorescence labeling in live cells and should become powerful tools for bioimaging applications.

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Language(s): eng - English
 Dates: 2013
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.2174/1381612811319300011
 Degree: -

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Title: Current Pharmaceutical Design
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 19 (30) Sequence Number: - Start / End Page: 5414 - 5420 Identifier: -