Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues
Chelatable, or mobile, forms of zinc play critical signaling roles in numerous biological processes. Elucidating the action of mobile Zn(II) in complex biological environments requires sensitive tools for visualizing, tracking, and manipulating Zn(II) ions. A large toolbox of synthetic photoinduced...
| Main Authors: | , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Chemical Society (ACS)
2016
|
| Online Access: | http://hdl.handle.net/1721.1/103921 https://orcid.org/0000-0002-9269-7815 https://orcid.org/0000-0002-2693-4982 https://orcid.org/0000-0002-5910-6948 |
| _version_ | 1826202246433472512 |
|---|---|
| author | Zastrow, Melissa L. Radford, Robert John Chyan, Wen Anderson, Charles T. Zhang, Daniel Y. Loas, Andrei Ioan Tzounopoulos, Thanos Lippard, Stephen J. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Zastrow, Melissa L. Radford, Robert John Chyan, Wen Anderson, Charles T. Zhang, Daniel Y. Loas, Andrei Ioan Tzounopoulos, Thanos Lippard, Stephen J. |
| author_sort | Zastrow, Melissa L. |
| collection | MIT |
| description | Chelatable, or mobile, forms of zinc play critical signaling roles in numerous biological processes. Elucidating the action of mobile Zn(II) in complex biological environments requires sensitive tools for visualizing, tracking, and manipulating Zn(II) ions. A large toolbox of synthetic photoinduced electron transfer (PET)-based fluorescent Zn(II) sensors are available, but the applicability of many of these probes is limited by poor zinc sensitivity and low dynamic ranges owing to proton interference. We present here a general approach for acetylating PET-based probes containing a variety of fluorophores and zinc-binding units. The new sensors provide substantially improved zinc sensitivity and allow for incubation of live cells and tissue slices with nM probe concentrations, a significant improvement compared to the μM concentrations that are typically required for a measurable fluorescence signal. Acetylation effectively reduces or completely quenches background fluorescence in the metal-free sensor. Binding of Zn(II) selectively and quickly mediates hydrolytic cleavage of the acetyl groups, providing a large fluorescence response. An acetylated blue coumarin-based sensor was used to carry out detailed analyses of metal binding and metal-promoted acetyl hydrolysis. Acetylated benzoresorufin-based red-emitting probes with different zinc-binding sites are effective for sensing Zn(II) ions in live cells when applied at low concentrations (∼50–100 nM). We used green diacetylated Zinpyr1 (DA-ZP1) to image endogenous mobile Zn(II) in the molecular layer of mouse dorsal cochlear nucleus (DCN), confirming that acetylation is a suitable approach for preparing sensors that are highly specific and sensitive to mobile zinc in biological systems. |
| first_indexed | 2024-09-23T12:04:29Z |
| format | Article |
| id | mit-1721.1/103921 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:04:29Z |
| publishDate | 2016 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1039212022-10-01T07:59:06Z Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues Zastrow, Melissa L. Radford, Robert John Chyan, Wen Anderson, Charles T. Zhang, Daniel Y. Loas, Andrei Ioan Tzounopoulos, Thanos Lippard, Stephen J. Massachusetts Institute of Technology. Department of Chemistry Zastrow, Melissa L. Radford, Robert John Chyan, Wen Zhang, Daniel Y. Loas, Andrei Ioan Lippard, Stephen J. Chelatable, or mobile, forms of zinc play critical signaling roles in numerous biological processes. Elucidating the action of mobile Zn(II) in complex biological environments requires sensitive tools for visualizing, tracking, and manipulating Zn(II) ions. A large toolbox of synthetic photoinduced electron transfer (PET)-based fluorescent Zn(II) sensors are available, but the applicability of many of these probes is limited by poor zinc sensitivity and low dynamic ranges owing to proton interference. We present here a general approach for acetylating PET-based probes containing a variety of fluorophores and zinc-binding units. The new sensors provide substantially improved zinc sensitivity and allow for incubation of live cells and tissue slices with nM probe concentrations, a significant improvement compared to the μM concentrations that are typically required for a measurable fluorescence signal. Acetylation effectively reduces or completely quenches background fluorescence in the metal-free sensor. Binding of Zn(II) selectively and quickly mediates hydrolytic cleavage of the acetyl groups, providing a large fluorescence response. An acetylated blue coumarin-based sensor was used to carry out detailed analyses of metal binding and metal-promoted acetyl hydrolysis. Acetylated benzoresorufin-based red-emitting probes with different zinc-binding sites are effective for sensing Zn(II) ions in live cells when applied at low concentrations (∼50–100 nM). We used green diacetylated Zinpyr1 (DA-ZP1) to image endogenous mobile Zn(II) in the molecular layer of mouse dorsal cochlear nucleus (DCN), confirming that acetylation is a suitable approach for preparing sensors that are highly specific and sensitive to mobile zinc in biological systems. National Institutes of Health (U.S.) (NIH grant GM065519) National Institutes of Health (U.S.) (NIH grant R01-DC007905) National Institutes of Health (U.S.) (NIH Fellowship (F32- EB019243)) National Institutes of Health (U.S.) (NIH Fellowship (T32-DC011499)) National Institutes of Health (U.S.) (NIH Fellowship (F32-DC013734)) 2016-08-15T20:03:28Z 2016-08-15T20:03:28Z 2015-09 2015-08 Article http://purl.org/eprint/type/JournalArticle 2379-3694 http://hdl.handle.net/1721.1/103921 Zastrow, Melissa L., Robert J. Radford, Wen Chyan, Charles T. Anderson, Daniel Y. Zhang, Andrei Loas, Thanos Tzounopoulos, and Stephen J. Lippard. "Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues." ACS Sensors 1:1 (2016), pp.32-39. © 2015 American Chemical Society. https://orcid.org/0000-0002-9269-7815 https://orcid.org/0000-0002-2693-4982 https://orcid.org/0000-0002-5910-6948 en_US http://dx.doi.org/10.1021/acssensors.5b00022 ACS Sensors 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) ACS |
| spellingShingle | Zastrow, Melissa L. Radford, Robert John Chyan, Wen Anderson, Charles T. Zhang, Daniel Y. Loas, Andrei Ioan Tzounopoulos, Thanos Lippard, Stephen J. Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues |
| title | Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues |
| title_full | Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues |
| title_fullStr | Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues |
| title_full_unstemmed | Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues |
| title_short | Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues |
| title_sort | reaction based probes for imaging mobile zinc in live cells and tissues |
| url | http://hdl.handle.net/1721.1/103921 https://orcid.org/0000-0002-9269-7815 https://orcid.org/0000-0002-2693-4982 https://orcid.org/0000-0002-5910-6948 |
| work_keys_str_mv | AT zastrowmelissal reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT radfordrobertjohn reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT chyanwen reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT andersoncharlest reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT zhangdaniely reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT loasandreiioan reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT tzounopoulosthanos reactionbasedprobesforimagingmobilezincinlivecellsandtissues AT lippardstephenj reactionbasedprobesforimagingmobilezincinlivecellsandtissues |