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Updated: Jun 3, 2025
Expression and Purification of Mammalian Bestrophin Ion Channels
Published on: August 2, 2018
Protocol for high-yield bacterial expression and purification of the voltage-dependent anion channel 1 for
Stefano Conti Nibali1, Andrea Magrì2, Angela Messina2
1Department of Molecular Medicine, University of Pavia, Pavia, Italy.
Abstract:
Voltage-dependent anion channel 1 (VDAC1) is a key protein in cellular metabolism and apoptosis. Here, we present a protocol to express and purify milligram amounts of recombinant VDAC1 in Escherichia coli. We detail steps for a fluorescence polarization-based high-throughput screening assay using NADH displacement, along with procedures for thermostability, fluorescence polarization, and X-ray crystallography. In this context, we demonstrate how 2-methyl-2,4-pentanediol (MPD), a crystallization reagent, interferes with VDAC1 small-molecule binding, hindering the detection of these ligands in the crystal. For complete details on the use and execution of this protocol, please refer to Conti Nibali et al.1.
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