Phasing and structure of bestrophin-1: a case study in the use of heavy-atom cluster compounds with multi-subunit transmembrane proteins.

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Authors
Kane Dickson, Veronica  ORCID logo  https://orcid.org/0000-0002-8649-8094
Abstract

The purification and three-dimensional crystallization of membrane proteins are commonly affected by a cumulation of pathologies that are less prevalent in their soluble counterparts. This may include severe anisotropy, poor spot shape, poor to moderate-resolution diffraction, crystal twinning, translational pseudo-symmetry and poor uptake of heavy atoms for derivatization. Such challenges must be circumvented by adaptations in the approach to crystallization and/or phasing. Here, an example of a protein that exhibited all of the above-mentioned complications is presented. Bestrophin-1 is a eukaryotic calcium-activated chloride channel, the structure of which was recently determined in complex with monoclonal antibody fragments using SAD phasing with tantalum bromide clusters (Ta6Br12·Br2). Some of the obstacles to obtaining improved diffraction and phasing for this particular channel are discussed, as well as the approach and adaptations that were key to determining the structure.

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Keywords
antibodies, bestrophin, detergents, membrane-protein crystallography, tantalum bromide, Animals, Bromides, Chickens, Chloride Channels, Crystallography, X-Ray, Models, Molecular, Protein Conformation, Protein Subunits, Tantalum
Journal Title
Acta Crystallogr D Struct Biol
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Journal ISSN
2059-7983
2059-7983
Volume Title
Publisher
International Union of Crystallography (IUCr)