Bestrophin interacts physically and functionally with protein phosphatase 2A

Lihua Y. Marmorstein, Precious J. McLaughlin, J. Brett Stanton, Lin Yan, John W. Crabb, Alan D. Marmorstein

Research output: Contribution to journalArticle

63 Scopus citations

Abstract

Bestrophin is a 68-kDa basolateral plasma membrane protein expressed in retinal pigment epithelial cells (RPE). It is encoded by the VMD2 gene, which is mutated in Best macular dystrophy, a disease characterized by a depressed light peak in the electrooculogram. Recently it was proposed that bestrophin is a chloride channel responsible for generating the light peak. To investigate its function further, we immunoaffinity purified a bestrophin complex from RPE lysates and identified bestrophin and the β-catalytic subunit of protein phosphatase 2A (PP2A) as members of the complex by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Protein-protein interaction between bestrophin and PP2Ac and the structural subunit of PP2A, PR65, was confirmed by reciprocal immunoprecipitation. The C-terminal cytoplasmic domain of bestrophin was sufficient for the interaction with PP2A as demonstrated by a pulldown assay using a fusion of this domain with glutathione S-transferase. Bestrophin was phosphorylated when expressed in RPE-J cells and this phosphorylation was sensitive to okadaic acid. Purified PP2A effectively dephosphorylated bestrophin in vitro. These data suggest that bestrophin is in the signal transduction pathway that modulates the light peak of the electrooculogram, that it is regulated by phosphorylation, and that phosphorylation of bestrophin is in turn regulated by PP2A.

Original languageEnglish (US)
Pages (from-to)30591-30597
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number34
DOIs
StatePublished - Aug 23 2002

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Marmorstein, L. Y., McLaughlin, P. J., Brett Stanton, J., Yan, L., Crabb, J. W., & Marmorstein, A. D. (2002). Bestrophin interacts physically and functionally with protein phosphatase 2A. Journal of Biological Chemistry, 277(34), 30591-30597. https://doi.org/10.1074/jbc.M204269200