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Bridging cells of three colors with two bio-orthogonal click reactions
Cell–cell interactions play a crucial role in the development and function of multicellular organisms. To study cell–cell interactions in vitro, it is a big challenge for researchers to artificially build up cell junctions to bridge different types of cells for this purpose. Herein, by employing two...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royal Society of Chemistry
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507188/ https://www.ncbi.nlm.nih.gov/pubmed/28757958 http://dx.doi.org/10.1039/c5sc01315a |
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author | Yuan, Yue Li, Di Zhang, Jia Chen, Xianmin Zhang, Chi Ding, Zhanling Wang, Lin Zhang, Xueqian Yuan, Junhua Li, Yinmei Kang, Yanbiao Liang, Gaolin |
author_facet | Yuan, Yue Li, Di Zhang, Jia Chen, Xianmin Zhang, Chi Ding, Zhanling Wang, Lin Zhang, Xueqian Yuan, Junhua Li, Yinmei Kang, Yanbiao Liang, Gaolin |
author_sort | Yuan, Yue |
collection | PubMed |
description | Cell–cell interactions play a crucial role in the development and function of multicellular organisms. To study cell–cell interactions in vitro, it is a big challenge for researchers to artificially build up cell junctions to bridge different types of cells for this purpose. Herein, by employing two orthogonal click reactions, we rationally designed four click reagents Mal-CBT, Mal-Cys, Mal-Alkyne, and Mal-N(3) and successfully applied them to bridge cells of three colors. Orthogonality between these two click reactions was validated in solution and characterized with HPLC and ESI-MS analyses. After modifications of fluorescent protein-expressing prokaryotic Escherichia coli (E. coli) cells (or eukaryotic HEK 293T cells) of three colors with the reagents Mal-Cys, Mal-CBT and Mal-Alkyne, or Mal-N(3), the cells were sequentially bridged. The HEK 293T cells showed a higher efficiency of cell bridging than the E. coli cells. Finally, using optical tweezers, we quantitatively measured the bridging probability between Mal-Cys-modified and Mal-CBT-modified HEK 293 cells, as well as the rupture force between two bridged cells. We found that the CBT–Cys click reaction markedly improved the efficiency of cell bridging and the rupture force between two bridged cells was measured to be 153.8 pN at a force-loading rate of 49 pN s(–1). Our results demonstrate that it is possible to use two (or n) orthogonal click reactions to bridge three (or n + 1) types of cells. Taking the biological importance of cell junctions into consideration, we anticipate that our method of bridging three types of cells with two bio-orthogonal click reactions will be a useful tool for biologists to study cell–cell interactions with more convenience and efficiency. |
format | Online Article Text |
id | pubmed-5507188 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-55071882017-07-28 Bridging cells of three colors with two bio-orthogonal click reactions Yuan, Yue Li, Di Zhang, Jia Chen, Xianmin Zhang, Chi Ding, Zhanling Wang, Lin Zhang, Xueqian Yuan, Junhua Li, Yinmei Kang, Yanbiao Liang, Gaolin Chem Sci Chemistry Cell–cell interactions play a crucial role in the development and function of multicellular organisms. To study cell–cell interactions in vitro, it is a big challenge for researchers to artificially build up cell junctions to bridge different types of cells for this purpose. Herein, by employing two orthogonal click reactions, we rationally designed four click reagents Mal-CBT, Mal-Cys, Mal-Alkyne, and Mal-N(3) and successfully applied them to bridge cells of three colors. Orthogonality between these two click reactions was validated in solution and characterized with HPLC and ESI-MS analyses. After modifications of fluorescent protein-expressing prokaryotic Escherichia coli (E. coli) cells (or eukaryotic HEK 293T cells) of three colors with the reagents Mal-Cys, Mal-CBT and Mal-Alkyne, or Mal-N(3), the cells were sequentially bridged. The HEK 293T cells showed a higher efficiency of cell bridging than the E. coli cells. Finally, using optical tweezers, we quantitatively measured the bridging probability between Mal-Cys-modified and Mal-CBT-modified HEK 293 cells, as well as the rupture force between two bridged cells. We found that the CBT–Cys click reaction markedly improved the efficiency of cell bridging and the rupture force between two bridged cells was measured to be 153.8 pN at a force-loading rate of 49 pN s(–1). Our results demonstrate that it is possible to use two (or n) orthogonal click reactions to bridge three (or n + 1) types of cells. Taking the biological importance of cell junctions into consideration, we anticipate that our method of bridging three types of cells with two bio-orthogonal click reactions will be a useful tool for biologists to study cell–cell interactions with more convenience and efficiency. Royal Society of Chemistry 2015-11-01 2015-07-28 /pmc/articles/PMC5507188/ /pubmed/28757958 http://dx.doi.org/10.1039/c5sc01315a Text en This journal is © The Royal Society of Chemistry 2015 http://creativecommons.org/licenses/by-nc/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported Licence (CC BY-NC 3.0) |
spellingShingle | Chemistry Yuan, Yue Li, Di Zhang, Jia Chen, Xianmin Zhang, Chi Ding, Zhanling Wang, Lin Zhang, Xueqian Yuan, Junhua Li, Yinmei Kang, Yanbiao Liang, Gaolin Bridging cells of three colors with two bio-orthogonal click reactions |
title | Bridging cells of three colors with two bio-orthogonal click reactions
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title_full | Bridging cells of three colors with two bio-orthogonal click reactions
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title_fullStr | Bridging cells of three colors with two bio-orthogonal click reactions
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title_full_unstemmed | Bridging cells of three colors with two bio-orthogonal click reactions
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title_short | Bridging cells of three colors with two bio-orthogonal click reactions
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title_sort | bridging cells of three colors with two bio-orthogonal click reactions |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507188/ https://www.ncbi.nlm.nih.gov/pubmed/28757958 http://dx.doi.org/10.1039/c5sc01315a |
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