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Photocrosslinking Approach to Investigate Protein Interactions in the BCL-2 Family

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BCL-2 Family Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1877))

Abstract

The Bcl-2 family of proteins regulates mitochondrial outer membrane permeability thereby making life or death decisions for cells. Most of Bcl-2 proteins contain hydrophobic regions that are embedded in intracellular membranes such as mitochondria. These membrane proteins are difficult to express and purify thereby preluding biochemical and biophysical characterizations. Here, we describe a photocrosslinking approach based on in vitro synthesis of Bcl-2 proteins with photoreactive amino acid analogs incorporated at specific locations. These photoreactive proteins are reconstituted into liposomal membranes with defined phospholipids or mitochondrial membranes isolated from animals, and their interactions with other Bcl-2 proteins are detected by photocrosslinking.

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Acknowledgments

This work was supported by the United States National Institutes of Health grants (R01GM062964, and P20GM103640), Oklahoma Center for the Advancement of Science and Technology grant (HR16-026), and Presbyterian Health Foundation grant (GRF00000125) to J.L.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Jialing Lin & Zhi Zhang

  2. Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Jialing Lin

  3. ReCode Therapeutics, Inc., Dallas, TX, USA

    Arthur E. Johnson

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  1. Jialing Lin
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  2. Arthur E. Johnson
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  3. Zhi Zhang
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Corresponding author

Correspondence to Jialing Lin .

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Editors and Affiliations

  1. Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY, USA

    Evripidis Gavathiotis

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Lin, J., Johnson, A.E., Zhang, Z. (2019). Photocrosslinking Approach to Investigate Protein Interactions in the BCL-2 Family. In: Gavathiotis, E. (eds) BCL-2 Family Proteins. Methods in Molecular Biology, vol 1877. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8861-7_9

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  • DOI: https://doi.org/10.1007/978-1-4939-8861-7_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8860-0

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