A role for myocilin in receptor-mediated endocytosis

Brian S. McKay, Nicole R. Congrove, Adiv A. Johnson, W. Michael Dismuke, Trent J. Bowen, W. Daniel Stamer

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Myocilin is a broadly expressed protein that when mutated uniquely causes glaucoma. While no function has been ascribed to explain focal disease, some properties of myocilin are known. Myocilin is a cytoplasmic protein that also localizes to vesicles specifically as part of a large membrane-associated complex with properties similar to the SNARE machinery that function in vesicle fusion. Its role in vesicle dynamics has not been detailed, however myocilin intersects with the endocytic compartment at the level of the multivesicular body. Since internalized GPCRs are sorted in the multivesicular body, we investigated whether myocilin functions in ligand-dependent GPR143 endocytosis. Using recombinant systems we found that the kinetics of myocilin recruitment to biotinylated membrane proteins was similar to that of arrestin-3. We also colocalized myocilin with GPR143 and Arrestin-2 by confocal microscopy. However, wild-type myocilin differed significantly in its association kinetics and co-localization with internalized proteins from mutant myocilin (P370L or T377M). Moreover, we found that myocilin bound to the cytoplasmic tail of GPR143, an interaction mediated by its amino terminal helix-turn-helix domain. Hydrodynamic analyses show that the myocilin-GPR143 protein complex is >158 kD and stable in 500 mM KCl, but not 0.1% SDS. Collectively, data indicate that myocilin is recruited to the membrane compartment, interacting with GPCR proteins during ligand-mediated endocytosis and that GPCR signaling underlies pathology in myocilin glaucoma.

Original languageEnglish (US)
Article numbere82301
JournalPloS one
Volume8
Issue number12
DOIs
StatePublished - Dec 18 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    McKay, B. S., Congrove, N. R., Johnson, A. A., Dismuke, W. M., Bowen, T. J., & Stamer, W. D. (2013). A role for myocilin in receptor-mediated endocytosis. PloS one, 8(12), [e82301]. https://doi.org/10.1371/journal.pone.0082301