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CW EPR and DEER Methods to Determine BCL-2 Family Protein Structure and Interactions: Application of Site-Directed Spin Labeling to BAK Apoptotic Pores

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Book cover BCL-2 Family Proteins

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

Abstract

The continuous wave (CW) and pulse electron paramagnetic resonance (EPR) methods enable the measurement of distances between spin-labeled residues in biopolymers including proteins, providing structural information. Here we describe the CW EPR deconvolution/convolution method and the four-pulse double electron–electron resonance (DEER) approach for distance determination, which were applied to elucidate the organization of the BAK apoptotic pores formed in the lipid bilayers.

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Acknowledgement

We thank Dr. Candice Klug at the National Biomedical EPR Center, Milwaukee, Wisconsin for the technical support in Q-band experiments. We also thank Drs. Gunnar Jeschke and Christian Altenbach for helpful discussions and Mr. Carter Grieve for his proofreading the manuscript. This work was supported by NIH grant R01GM097508, EPR Center of the Rosalind Franklin University of Medicine and Science, and the RFUMS Start-up fund to. K.J.O.; NIH grant P01 GM080513 to E.J.H.; NIH grants RR022422 (DEER instrumentation grant), OD011937 (DEER instrumentation grant), and EB001980 (National Biomedical EPR Center grant) to C.K; and NSF Cooperative Agreement No. DMR-1644779 to L.S.

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Authors and Affiliations

  1. Department of Biochemistry & Molecular Biology, The Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA

    Tirtha Mandal & Kyoung Joon Oh

  2. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA

    Tirtha Mandal

  3. Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN, USA

    Eric J. Hustedt

  4. National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL, USA

    Likai Song

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  1. Tirtha Mandal
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  2. Eric J. Hustedt
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  3. Likai Song
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  4. Kyoung Joon Oh
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Corresponding author

Correspondence to Kyoung Joon Oh .

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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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Mandal, T., Hustedt, E.J., Song, L., Oh, K.J. (2019). CW EPR and DEER Methods to Determine BCL-2 Family Protein Structure and Interactions: Application of Site-Directed Spin Labeling to BAK Apoptotic Pores. 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_18

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  • DOI: https://doi.org/10.1007/978-1-4939-8861-7_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8860-0

  • Online ISBN: 978-1-4939-8861-7

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