Jump to main content
Jump to site search

Issue 2, 2015
Previous Article Next Article

A novel copper-chelating strategy for fluorescent proteins to image dynamic copper fluctuations on live cell surfaces

Author affiliations

Abstract

Copper is indispensable in most aerobic organisms although it is toxic if unregulated as illustrated in many neurodegenerative diseases. To elucidate the mechanisms underlying copper release from cells, a membrane-targeting reporter which can compete with extracellular copper-binding molecules is highly desirable. However, engineering a reporter protein to provide both high sensitivity and selectivity for copper(II) has been challenging, likely due to a lack of proper copper(II)-chelating strategies within proteins. Here, we report a new genetically encoded fluorescent copper(II) reporter by employing a copper-binding tripeptide derived from human serum albumin (HSA), which is one of the major copper-binding proteins in extracellular environments. Optimized insertion of the tripeptide into the green fluorescent protein leads to rapid fluorescence quenching (up to >85% change) upon copper-binding, while other metal ions have no effect. Furthermore, the high binding affinity of the reporter enables reliable copper detection even in the presence of competing biomolecules such as HSA and amyloid beta peptides. We also demonstrate that our reporter proteins can be used to visualize dynamic copper fluctuations on living HeLa cell surfaces.

Graphical abstract: A novel copper-chelating strategy for fluorescent proteins to image dynamic copper fluctuations on live cell surfaces

Back to tab navigation

Supplementary files

Article information


Submitted
02 Oct 2014
Accepted
18 Nov 2014
First published
19 Nov 2014

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2015,6, 1301-1307
Article type
Edge Article

A novel copper-chelating strategy for fluorescent proteins to image dynamic copper fluctuations on live cell surfaces

Y. Choi, J. O. Keem, C. Y. Kim, H. R. Yoon, W. D. Heo, B. H. Chung and Y. Jung, Chem. Sci., 2015, 6, 1301
DOI: 10.1039/C4SC03027C

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material.

Reproduced material should be attributed as follows:

  • For reproduction of material from NJC:
    [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
  • For reproduction of material from PCCP:
    [Original citation] - Published by the PCCP Owner Societies.
  • For reproduction of material from PPS:
    [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
  • For reproduction of material from all other RSC journals:
    [Original citation] - Published by The Royal Society of Chemistry.

Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.


Social activity

Search articles by author

Spotlight

Advertisements