A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells
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nihms-1012386.pdf
Description
Accepted version
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1.12 MB
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Author(s)
Richardson, Christopher E. R.
Nolan, Elizabeth Marie
Shoulders, Matthew D.
Lippard, Stephen J.
Date Issued
December 2018
Journal
Biochemistry
Publisher
American Chemical Society (ACS)
Citation
Richardson, Christopher E. R. et al. “A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells.” Biochemistry 57 (2018): 6807-6815 © 2018 The Author(s)
Version
Author's final manuscript
Abstract
Sensitive measurements of cellular Zn(II) uptake currently rely on quantitating radioactive emissions from cells treated with 65 Zn(II). Here, we describe a straightforward and reliable method employing a stable isotope to sensitively measure Zn(II) uptake by metazoan cells. First, biological medium selectively depleted of natural abundance Zn(II) using A12-resin [Richardson, C. E. R., et al. (2018) J. Am. Chem. Soc. 140, 2413] is restored to physiological levels of Zn(II) by addition of a non-natural Zn(II) isotope distribution comprising 70% 70 Zn(II). The resulting 70 Zn(II)-enriched medium facilitates quantitation of Zn(II) uptake using inductively coupled plasma-mass spectrometry (ICP-MS). This sensitive and reliable assay assesses Zn(II)-uptake kinetics at early time points and can be used to delineate how chemical and genetic perturbations influence Zn(II) uptake. Further, the use of ICP-MS in a Zn(II)-uptake assay permits simultaneous measurement of multiple metal ion concentrations. We used this capability to show that, across three cell lines, Zn(II) deficiency enhances selectivity for Zn(II) over Cd(II) uptake.
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/ACS.BIOCHEM.8B01043
