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Engineered Cross-Linking to Study the Pore Architecture of the CRAC Channel

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The CRAC Channel

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1843))

Abstract

ORAI1 constitutes the pore-forming subunit of the calcium release-activated calcium (CRAC) channel, a prototypical store-operated channel that is essential for the activation of cells of the immune system. Here we describe a convenient yet powerful cross-linking approach to examine the pore architecture of CRAC channels using ORAI1 proteins engineered to contain one or two cysteine residues. The generalizable cross-linking in situ approach can also be readily extended to study other integral membrane proteins expressed in various types of cells.

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Acknowledgments

We thank the financial supports from the National Institutes of Health grant (R01GM112003, R21GM126532 and R01HL134780), the Welch Foundation (BE-1913), the American Cancer Society (RSG-16-215-01 TBE), the Cancer Prevention and Research Institute of Texas (RR140053), the American Heart Association (16IRG27250155), and the John S. Dunn Foundation and by an allocation from the Texas A&M University Health Science Center Startup Fund.

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Authors and Affiliations

  1. Center for Translational Cancer Research, Institute of Biosciences and Technology, College of Medicine, Texas A&M University, 2121 W. Holcombe Blvd, Houston, TX, USA

    Guolin Ma, Lian He, Ji Jing, Peng Tan, Yun Huang & Yubin Zhou

  2. Department of Medical Physiology, College of Medicine, Texas A&M University, Temple, TX, USA

    Yubin Zhou

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  1. Guolin Ma
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  2. Lian He
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  3. Ji Jing
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  4. Peng Tan
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  5. Yun Huang
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  6. Yubin Zhou
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Corresponding author

Correspondence to Yubin Zhou .

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Editors and Affiliations

  1. IRSET Inserm UMR 1085/ UR1, Rennes, France

    Aubin Penna

  2. CNRS ERL 7003, Laboratoire STIM, Poitiers, France

    Bruno Constantin

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Ma, G., He, L., Jing, J., Tan, P., Huang, Y., Zhou, Y. (2018). Engineered Cross-Linking to Study the Pore Architecture of the CRAC Channel. In: Penna, A., Constantin, B. (eds) The CRAC Channel. Methods in Molecular Biology, vol 1843. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8704-7_13

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  • DOI: https://doi.org/10.1007/978-1-4939-8704-7_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8702-3

  • Online ISBN: 978-1-4939-8704-7

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