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Photoactivatable and photoconvertible fluorescent probes for protein labeling.
ACS Chem Biol. 2010 May 21; 5(5):507-16.AC

Abstract

Photosensitive probes are powerful tools to study cellular processes with high temporal and spatial resolution. However, most synthetic fluorophores suited for biomolecular imaging have not been converted yet to appropriate photosensitive analogues. Here we describe a generally applicable strategy for the generation of photoactivatable and photoconvertible fluorescent probes that can be selectively coupled to SNAP-tag fusion proteins in living cells. Photoactivatable versions of fluorescein and Cy3 as well as a photoconvertible Cy5-Cy3 probe were prepared and coupled to selected proteins on the cell surface, in the cytosol, and in the nucleus of cells. In proof-of-principle experiments, the photoactivatable Cy3 probe was used to characterize the mobility of a lipid-anchored cell surface protein and of a G protein coupled receptor (GPCR). This work establishes a generally applicable strategy for the generation of a large variety of different photosensitive fluorophores with tailor-made properties for biomolecular imaging.

Authors+Show Affiliations

Institute of Chemical Sciences and Engineering, Ecole Polytechnique Federale de Lausanne, Switzerland.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20218675

Citation

Maurel, Damien, et al. "Photoactivatable and Photoconvertible Fluorescent Probes for Protein Labeling." ACS Chemical Biology, vol. 5, no. 5, 2010, pp. 507-16.
Maurel D, Banala S, Laroche T, et al. Photoactivatable and photoconvertible fluorescent probes for protein labeling. ACS Chem Biol. 2010;5(5):507-16.
Maurel, D., Banala, S., Laroche, T., & Johnsson, K. (2010). Photoactivatable and photoconvertible fluorescent probes for protein labeling. ACS Chemical Biology, 5(5), 507-16. https://doi.org/10.1021/cb1000229
Maurel D, et al. Photoactivatable and Photoconvertible Fluorescent Probes for Protein Labeling. ACS Chem Biol. 2010 May 21;5(5):507-16. PubMed PMID: 20218675.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Photoactivatable and photoconvertible fluorescent probes for protein labeling. AU - Maurel,Damien, AU - Banala,Sambashiva, AU - Laroche,Thierry, AU - Johnsson,Kai, PY - 2010/3/12/entrez PY - 2010/3/12/pubmed PY - 2010/8/27/medline SP - 507 EP - 16 JF - ACS chemical biology JO - ACS Chem. Biol. VL - 5 IS - 5 N2 - Photosensitive probes are powerful tools to study cellular processes with high temporal and spatial resolution. However, most synthetic fluorophores suited for biomolecular imaging have not been converted yet to appropriate photosensitive analogues. Here we describe a generally applicable strategy for the generation of photoactivatable and photoconvertible fluorescent probes that can be selectively coupled to SNAP-tag fusion proteins in living cells. Photoactivatable versions of fluorescein and Cy3 as well as a photoconvertible Cy5-Cy3 probe were prepared and coupled to selected proteins on the cell surface, in the cytosol, and in the nucleus of cells. In proof-of-principle experiments, the photoactivatable Cy3 probe was used to characterize the mobility of a lipid-anchored cell surface protein and of a G protein coupled receptor (GPCR). This work establishes a generally applicable strategy for the generation of a large variety of different photosensitive fluorophores with tailor-made properties for biomolecular imaging. SN - 1554-8937 UR - https://www.unboundmedicine.com/medline/citation/20218675/Photoactivatable_and_photoconvertible_fluorescent_probes_for_protein_labeling_ L2 - https://dx.doi.org/10.1021/cb1000229 DB - PRIME DP - Unbound Medicine ER -