Peptide-Based, Two-Fluorophore, Ratiometric Probe for Quantifying Mobile Zinc in Biological Solutions
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LIppard_Peptide-based.pdf
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Author(s)
Azrad, Maria
Demark-Wahnefried, Wendy
Frederickson, Christopher J.
Zhang, Daniel Y.
Lippard, Stephen J.
Radford, Robert John
Date Issued
November 2014
Journal
ACS Chemical Biology
Publisher
American Chemical Society (ACS)
Citation
Zhang, Daniel Y.; Azrad, Maria; Demark-Wahnefried, Wendy; Frederickson, Christopher J.; Lippard, Stephen J. and Radford, Robert J. "Peptide-Based, Two-Fluorophore, Ratiometric Probe for Quantifying Mobile Zinc in Biological Solutions." ACS Chemical Biology 10, 2 (February 2015): 385–389 © 2014 American Chemical Society
Version
Final published version
Abstract
Small-molecule fluorescent sensors are versatile agents for detecting mobile zinc in biology. Capitalizing on the abundance of validated mobile zinc probes, we devised a strategy for repurposing existing intensity-based sensors for quantitative applications. Using solid-phase peptide synthesis, we conjugated a zinc-sensitive Zinpyr-1 derivative and a zinc-insensitive 7-hydroxycoumarin derivative onto opposite ends of a rigid P₉K peptide scaffold to create HcZ9, a ratiometric fluorescent probe for mobile zinc. A plate reader-based assay using HcZ9 was developed, the accuracy of which is comparable to that of atomic absorption spectroscopy. We investigated zinc accumulation in prostatic cells and zinc levels in human seminal fluid. When normal and tumorigenic cells are bathed in zinc-enriched media, cellular mobile zinc is buffered and changes slightly, but total zinc levels increase significantly. Quantification of mobile and total zinc levels in human seminal plasma revealed that the two are positively correlated with a Pearson’s coefficient of 0.73.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1021/cb500617c
