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Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays
Bcl-2 family proteins have important roles in tumor initiation, progression and resistance to therapy. Pro-survival Bcl-2 proteins are regulated by their interactions with pro-death BH3-only proteins making these protein–protein interactions attractive therapeutic targets. Although these interaction...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5424096/ https://www.ncbi.nlm.nih.gov/pubmed/22460384 http://dx.doi.org/10.1038/cddis.2012.28 |
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author | Wong, C Anderson, D J Lee, E F Fairlie, W D Ludlam, M J C |
author_facet | Wong, C Anderson, D J Lee, E F Fairlie, W D Ludlam, M J C |
author_sort | Wong, C |
collection | PubMed |
description | Bcl-2 family proteins have important roles in tumor initiation, progression and resistance to therapy. Pro-survival Bcl-2 proteins are regulated by their interactions with pro-death BH3-only proteins making these protein–protein interactions attractive therapeutic targets. Although these interactions have been extensively characterized biochemically, there is a paucity of tools to assess these interactions in cells. Here, we address this limitation by developing quantitative, high throughput microscopy assays to characterize Bcl-2 and BH3-only protein interactions in live cells. We use fluorescent proteins to label the interacting proteins of interest, enabling visualization and quantification of their mitochondria-localized interactions. Using tool compounds, we demonstrate the suitability of our assays to characterize the cellular activity of putative therapeutic molecules that target the interaction between pro-survival Bcl-2 and pro-death BH3-only proteins. In addition to the relevance of our assays for drug discovery, we anticipate that our work will contribute to an improved understanding of the mechanisms that regulate these important protein–protein interactions within the cell. |
format | Online Article Text |
id | pubmed-5424096 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-54240962017-05-19 Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays Wong, C Anderson, D J Lee, E F Fairlie, W D Ludlam, M J C Cell Death Dis Original Article Bcl-2 family proteins have important roles in tumor initiation, progression and resistance to therapy. Pro-survival Bcl-2 proteins are regulated by their interactions with pro-death BH3-only proteins making these protein–protein interactions attractive therapeutic targets. Although these interactions have been extensively characterized biochemically, there is a paucity of tools to assess these interactions in cells. Here, we address this limitation by developing quantitative, high throughput microscopy assays to characterize Bcl-2 and BH3-only protein interactions in live cells. We use fluorescent proteins to label the interacting proteins of interest, enabling visualization and quantification of their mitochondria-localized interactions. Using tool compounds, we demonstrate the suitability of our assays to characterize the cellular activity of putative therapeutic molecules that target the interaction between pro-survival Bcl-2 and pro-death BH3-only proteins. In addition to the relevance of our assays for drug discovery, we anticipate that our work will contribute to an improved understanding of the mechanisms that regulate these important protein–protein interactions within the cell. Nature Publishing Group 2012-03 2012-03-29 /pmc/articles/PMC5424096/ /pubmed/22460384 http://dx.doi.org/10.1038/cddis.2012.28 Text en Copyright © 2012 Macmillan Publishers Limited http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Original Article Wong, C Anderson, D J Lee, E F Fairlie, W D Ludlam, M J C Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
title | Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
title_full | Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
title_fullStr | Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
title_full_unstemmed | Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
title_short | Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
title_sort | direct visualization of bcl-2 family protein interactions using live cell fluorescent protein redistribution assays |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5424096/ https://www.ncbi.nlm.nih.gov/pubmed/22460384 http://dx.doi.org/10.1038/cddis.2012.28 |
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