YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C
Genetically encoded calcium indicators based on truncated troponin C are attractive probes for calcium imaging due to their relatively small molecular size and twofold reduced calcium ion buffering. However, the best‐suited members of this family, YTnC and cNTnC, suffer from low molecular brightness...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2023-11-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.13702 |
| _version_ | 1797635880289042432 |
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| author | Oksana M. Subach Anna V. Vlaskina Yulia K. Agapova Alena Y. Nikolaeva Anna M. Varizhuk Oleg V. Podgorny Kiryl D. Piatkevich Maxim V. Patrushev Konstantin M. Boyko Fedor V. Subach |
| author_facet | Oksana M. Subach Anna V. Vlaskina Yulia K. Agapova Alena Y. Nikolaeva Anna M. Varizhuk Oleg V. Podgorny Kiryl D. Piatkevich Maxim V. Patrushev Konstantin M. Boyko Fedor V. Subach |
| author_sort | Oksana M. Subach |
| collection | DOAJ |
| description | Genetically encoded calcium indicators based on truncated troponin C are attractive probes for calcium imaging due to their relatively small molecular size and twofold reduced calcium ion buffering. However, the best‐suited members of this family, YTnC and cNTnC, suffer from low molecular brightness, limited dynamic range, and/or poor sensitivity to calcium transients in neurons. To overcome these limitations, we developed an enhanced version of YTnC, named YTnC2. Compared with YTnC, YTnC2 had 5.7‐fold higher molecular brightness and 6.4‐fold increased dynamic range in vitro. YTnC2 was successfully used to reveal calcium transients in the cytosol and in the lumen of mitochondria of both mammalian cells and cultured neurons. Finally, we obtained and analyzed the crystal structure of the fluorescent domain of the YTnC2 mutant. |
| first_indexed | 2024-03-11T12:27:22Z |
| format | Article |
| id | doaj.art-1f1660325fa647b5af79a6e6567d9c71 |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-03-11T12:27:22Z |
| publishDate | 2023-11-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-1f1660325fa647b5af79a6e6567d9c712023-11-06T07:27:07ZengWileyFEBS Open Bio2211-54632023-11-0113112047206010.1002/2211-5463.13702YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin COksana M. Subach0Anna V. Vlaskina1Yulia K. Agapova2Alena Y. Nikolaeva3Anna M. Varizhuk4Oleg V. Podgorny5Kiryl D. Piatkevich6Maxim V. Patrushev7Konstantin M. Boyko8Fedor V. Subach9Complex of NBICS Technologies National Research Center “Kurchatov Institute” Moscow RussiaComplex of NBICS Technologies National Research Center “Kurchatov Institute” Moscow RussiaComplex of NBICS Technologies National Research Center “Kurchatov Institute” Moscow RussiaComplex of NBICS Technologies National Research Center “Kurchatov Institute” Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine of Federal Medical Biological Agency Moscow RussiaM.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry RAS Moscow RussiaSchool of Life Sciences Westlake University Hangzhou ChinaComplex of NBICS Technologies National Research Center “Kurchatov Institute” Moscow RussiaBach Institute of Biochemistry Research Centre of Biotechnology of the Russian Academy of Sciences Moscow RussiaComplex of NBICS Technologies National Research Center “Kurchatov Institute” Moscow RussiaGenetically encoded calcium indicators based on truncated troponin C are attractive probes for calcium imaging due to their relatively small molecular size and twofold reduced calcium ion buffering. However, the best‐suited members of this family, YTnC and cNTnC, suffer from low molecular brightness, limited dynamic range, and/or poor sensitivity to calcium transients in neurons. To overcome these limitations, we developed an enhanced version of YTnC, named YTnC2. Compared with YTnC, YTnC2 had 5.7‐fold higher molecular brightness and 6.4‐fold increased dynamic range in vitro. YTnC2 was successfully used to reveal calcium transients in the cytosol and in the lumen of mitochondria of both mammalian cells and cultured neurons. Finally, we obtained and analyzed the crystal structure of the fluorescent domain of the YTnC2 mutant.https://doi.org/10.1002/2211-5463.13702calcium imagingcrystal structuregenetically encoded calcium indicatorprotein engineeringtroponin CYTnC2 |
| spellingShingle | Oksana M. Subach Anna V. Vlaskina Yulia K. Agapova Alena Y. Nikolaeva Anna M. Varizhuk Oleg V. Podgorny Kiryl D. Piatkevich Maxim V. Patrushev Konstantin M. Boyko Fedor V. Subach YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C FEBS Open Bio calcium imaging crystal structure genetically encoded calcium indicator protein engineering troponin C YTnC2 |
| title | YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C |
| title_full | YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C |
| title_fullStr | YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C |
| title_full_unstemmed | YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C |
| title_short | YTnC2, an improved genetically encoded green calcium indicator based on toadfish troponin C |
| title_sort | ytnc2 an improved genetically encoded green calcium indicator based on toadfish troponin c |
| topic | calcium imaging crystal structure genetically encoded calcium indicator protein engineering troponin C YTnC2 |
| url | https://doi.org/10.1002/2211-5463.13702 |
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