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

  • Guolin Ma
  • Lian He
  • Ji Jing
  • Peng Tan
  • Yun Huang
  • Yubin ZhouEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1843)


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.

Key words

Calcium signaling ORAI1 Calcium release-activated calcium channel Structure Disulfide bond Cross-linking Membrane protein Lymphocyte activation Stromal interaction molecule Store-operated calcium entry 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Guolin Ma
    • 1
  • Lian He
    • 1
  • Ji Jing
    • 1
  • Peng Tan
    • 1
  • Yun Huang
    • 1
  • Yubin Zhou
    • 1
    • 2
    Email author
  1. 1.Center for Translational Cancer Research, Institute of Biosciences and Technology, College of Medicine Texas A&M UniversityHoustonUSA
  2. 2.Department of Medical Physiology, College of MedicineTexas A&M UniversityTempleUSA

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