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Functional assembly and purinergic activation of bestrophins

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Abstract

Proteins of the bestrophin family produce Ca2+-activated Cl− currents and regulate voltage-gated Ca2+ channels. Bestrophin 1 was first identified in the retinal pigment epithelium. Four human paralogs (hBest1–hBest4) exist, and for some bestrophins, dimeric and heterotetrameric structures have been proposed. Here, we demonstrate that hBest1–hBest4 induce Cl− conductances of different amplitudes when expressed in HEK293 cells and when activated through purinergic stimulation. hBest1 mutants that are known to cause autosomal dominant macular dystrophy (Best disease) did not produce a Cl− current. Bestrophins were colocalized and showed molecular and functional interaction in HEK293 cells, overexpressing hBest1 and hBest2 or hBest4. Interaction was confirmed in airway epithelial cells coexpressing endogenous bestrophins. A fraction of hBest2 and hBest4 was expressed in the membrane, while most of hBest1 was found in the endoplasmic reticulum. Nevertheless, hBest1 has a clear role for the adenosine triphosphate (ATP; or uridine triphosphate)-induced Cl− current in both HEK293 and Calu-3 cells. Since native epithelial tissues typically express several bestrophin paralogs, these proteins may exist as heterooligomeric structures.

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Acknowledgments

DFG SFB699, DFG KU 756/8-2 and Else Kröner-Fresenius-Stiftung P36/05//A44/05. We gratefully acknowledge the supply of human Best-cDNA’s by Dr. J. Nathans, Department of Neuroscience, Johns Hopkins University, Baltimore. We acknowledge the expert technical assistance by Ms. A. Paech. The authors thank Dr. Miriam Breunig (Pharmazeutische Technologie, Universität Regensburg) for providing access to the confocal microscope.

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  1. Institut für Physiologie, Universität Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany

    Vladimir M. Milenkovic, René Barro Soria, Fadi Aldehni, Rainer Schreiber & Karl Kunzelmann

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  1. Vladimir M. Milenkovic
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  2. René Barro Soria
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Correspondence to Karl Kunzelmann.

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Milenkovic, V.M., Soria, R.B., Aldehni, F. et al. Functional assembly and purinergic activation of bestrophins. Pflugers Arch - Eur J Physiol 458, 431–441 (2009). https://doi.org/10.1007/s00424-008-0626-y

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  • DOI: https://doi.org/10.1007/s00424-008-0626-y

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