NDLI: Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Wong, C. Anderson, D. J. Lee, E. F. Fairlie, W. D. Ludlam, M. J. C.
Description	Country affiliation: United States Author Affiliation: Wong C ( Department of Research Oncology, Genentech, Inc., South San Francisco, CA 90480, USA.)
Abstract	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.
File Format	HTM / HTML
e-ISSN	20414889
Journal	Cell Death and Disease
Volume Number	3
Language	English
Publisher	Nature Publishing Group
Publisher Date	2012-03-29
Publisher Place	Great Britain (UK)
Access Restriction	Open
Subject Keyword	Research Support, Non-u.s. Gov't Bh3 Interacting Domain Death Agonist Protein Metabolism Recombinant Fusion Proteins Protein Interaction Mapping Discipline Cell Biology High-throughput Screening Assays Mitochondria Hek293 Cells Genetics Proto-oncogene Proteins C-bcl-2 Luminescent Proteins Apoptosis
Content Type	Text
Resource Type	Article

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