5T5N

Calcium-activated chloride channel bestrophin-1 (BEST1), triple mutant: I76A, F80A, F84A; in complex with an Fab antibody fragment, chloride, and calcium

Summary for 5T5N

• Entry DOI: 10.2210/pdb5t5n/pdb
• Descriptor: bestrophin-1 (BEST1), Fab antibody fragment, light chain (10D10), Fab antibody fragment, heavy chain (10D10), ... (8 entities in total)
• Functional Keywords: ion channel, anion, chloride, calcium-activated, eukaryotic membrane protein, cacc, membrane protein
• Biological source: Gallus gallus (Chicken); More
• Total number of polymer chains: 15
• Total formula weight: 473379.65

Authors

Long, S.B.,Vaisey, G. (deposition date: 2016-08-31, release date: 2016-11-30, Last modification date: 2024-11-20)

Primary citation

Vaisey, G.,Miller, A.N.,Long, S.B.
Distinct regions that control ion selectivity and calcium-dependent activation in the bestrophin ion channel.
Proc. Natl. Acad. Sci. U.S.A., 113:E7399-E7408, 2016

PubMed Abstract: Cytoplasmic calcium (Ca) activates the bestrophin anion channel, allowing chloride ions to flow down their electrochemical gradient. Mutations in bestrophin 1 (BEST1) cause macular degenerative disorders. Previously, we determined an X-ray structure of chicken BEST1 that revealed the architecture of the channel. Here, we present electrophysiological studies of purified wild-type and mutant BEST1 channels and an X-ray structure of a Ca-independent mutant. From these experiments, we identify regions of BEST1 responsible for Ca activation and ion selectivity. A "Ca clasp" within the channel's intracellular region acts as a sensor of cytoplasmic Ca. Alanine substitutions within a hydrophobic "neck" of the pore, which widen it, cause the channel to be constitutively active, irrespective of Ca. We conclude that the primary function of the neck is as a "gate" that controls chloride permeation in a Ca-dependent manner. In contrast to what others have proposed, we find that the neck is not a major contributor to the channel's ion selectivity. We find that mutation of a cytosolic "aperture" of the pore does not perturb the Ca dependence of the channel or its preference for anions over cations, but its mutation dramatically alters relative permeabilities among anions. The data suggest that the aperture functions as a size-selective filter that permits the passage of small entities such as partially dehydrated chloride ions while excluding larger molecules such as amino acids. Thus, unlike ion channels that have a single "selectivity filter," in bestrophin, distinct regions of the pore govern anion-vs.-cation selectivity and the relative permeabilities among anions.

PubMed: 27821745
DOI: 10.1073/pnas.1614688113

Experimental method

X-RAY DIFFRACTION (3.1 Å)