Engineered Cross-Linking to Study the Pore Architecture of the CRAC Channel
Guolin Ma1, Lian He1, Ji Jing1
1Center for Translational Cancer Research, Institute of Biosciences and Technology, College of Medicine, Texas A&M University, 2121 W. Holcombe Blvd, Houston, TX, USA.
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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