A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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American Chemical Society (ACS)
2020
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| Online Access: | https://hdl.handle.net/1721.1/125596 |
| _version_ | 1826214787412918272 |
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| author | Richardson, Christopher E. R. Nolan, Elizabeth Marie Shoulders, Matthew D. Lippard, Stephen J. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Richardson, Christopher E. R. Nolan, Elizabeth Marie Shoulders, Matthew D. Lippard, Stephen J. |
| author_sort | Richardson, Christopher E. R. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T16:11:08Z |
| format | Article |
| id | mit-1721.1/125596 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T16:11:08Z |
| publishDate | 2020 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1255962022-09-29T18:49:39Z A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells Richardson, Christopher E. R. Nolan, Elizabeth Marie Shoulders, Matthew D. Lippard, Stephen J. Massachusetts Institute of Technology. Department of Chemistry 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. National Institute of Environmental Health Sciences (Grant P30-ES002109) National Institute of General Medical Sciences (U.S.) (Grant R01-GM065519) National Institute of Biomedical Imaging and Bioengineering (Grant T32-EB019940) National Cancer Institute (U.S.) (Grant P30-CA14051) 2020-06-01T16:41:47Z 2020-06-01T16:41:47Z 2018-12 2020-01-02T16:31:01Z Article http://purl.org/eprint/type/JournalArticle 0006-2960 https://hdl.handle.net/1721.1/125596 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) en https://dx.doi.org/10.1021/ACS.BIOCHEM.8B01043 Biochemistry 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. application/pdf American Chemical Society (ACS) PMC |
| spellingShingle | Richardson, Christopher E. R. Nolan, Elizabeth Marie Shoulders, Matthew D. Lippard, Stephen J. A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells |
| title | A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells |
| title_full | A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells |
| title_fullStr | A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells |
| title_full_unstemmed | A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells |
| title_short | A Sensitive, Nonradioactive Assay for Zn(II) Uptake into Metazoan Cells |
| title_sort | sensitive nonradioactive assay for zn ii uptake into metazoan cells |
| url | https://hdl.handle.net/1721.1/125596 |
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