Jump to main content
Jump to site search

Issue 7, 2015
Previous Article Next Article

Near-instant surface-selective fluorogenic protein quantification using sulfonated triarylmethane dyes and fluorogen activating proteins

Author affiliations

Abstract

Agonist-promoted G-protein coupled receptor (GPCR) endocytosis and recycling plays an important role in many signaling events in the cell. However, the approaches that allow fast and quantitative analysis of such processes still remain limited. Here we report an improved labeling approach based on the genetic fusion of a fluorogen activating protein (FAP) to a GPCR and binding of a sulfonated analog of the malachite green (MG) fluorogen to rapidly and selectively label cell surface receptors. Fluorescence microscopy and flow cytometry demonstrate that this dye does not cross the plasma membrane, binds with high affinity to a dL5** FAP-GPCR fusion construct, activating tagged surface receptors within seconds of addition. The ability to rapidly and selectively label cell surface receptors with a fluorogenic genetically encoded tag allows quantitative imaging and analysis of highly dynamic processes like receptor endocytosis and recycling.

Graphical abstract: Near-instant surface-selective fluorogenic protein quantification using sulfonated triarylmethane dyes and fluorogen activating proteins

Back to tab navigation

Supplementary files

Publication details

The article was received on 30 Oct 2014, accepted on 09 Dec 2014 and first published on 10 Dec 2014


Article type: Paper
DOI: 10.1039/C4OB02309A
Author version
available:
Download author version (PDF)
Org. Biomol. Chem., 2015,13, 2078-2086

  •   Request permissions

    Near-instant surface-selective fluorogenic protein quantification using sulfonated triarylmethane dyes and fluorogen activating proteins

    Q. Yan, B. F. Schmidt, L. A. Perkins, M. Naganbabu, S. Saurabh, S. K. Andreko and M. P. Bruchez, Org. Biomol. Chem., 2015, 13, 2078
    DOI: 10.1039/C4OB02309A

Search articles by author

Spotlight

Advertisements