Issue 7, 2019

A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede

Abstract

Self-labelling protein tags with fluorogenic probes serve as great fluorescence imaging tools to understand key questions of protein dynamics and functions in living cells. In the present study, we report a SNAP-tag fluorogenic probe 4c mimicking the chromophore of the red fluorescent protein Kaede. The molecular rotor properties of 4c were utilized as a fluorogenic probe for SNAP-tag, such that conjugation with SNAPf protein leads to inhibition of twisted intramolecular charge transfer, triggering fluorogenecity. Upon conjugation with SNAPf, 4c exhibited approximately a 90-fold enhancement in fluorescence intensity with fast labelling kinetics (k2 = 15 000 M−1 s−1). Biochemical and spectroscopic studies confirmed that fluorescence requires formation of folded SNAPf·4c covalent conjugate between Cys 145 and 4c. Confocal microscopy and flow cytometry showed that 4c is capable of detecting SNAPf proteins or SNAPf fused with a protein of interest in living cells. This work provides a framework to develop the large family of FP chromophores into fluorogenic probes for self-labelling protein tags.

Graphical abstract: A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede

Supplementary files

Article information

Article type
Paper
Submitted
22 जून 2018
Accepted
25 सितम्बर 2018
First published
25 सितम्बर 2018

Org. Biomol. Chem., 2019,17, 1906-1915

A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede

K. H. Jung, M. Fares, L. S. Grainger, C. H. Wolstenholme, A. Hou, Y. Liu and X. Zhang, Org. Biomol. Chem., 2019, 17, 1906 DOI: 10.1039/C8OB01483C

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements