Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration

John D. Hulleman, Shalesh Kaushal, William E. Balch, Jeffery W. Kelly

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

An Arg345Trp (R345W) mutation in epidermal growth factor-containing, fibulin-like extracellular matrix protein 1 (EFEMP1) causes its inefficient secretion and the macular dystrophy malattia leventinese/Doyne honeycomb retinal dystrophy (ML/DHRD). To understand the influence of the protein homeostasis (or proteostasis) network in rescuing mutant EFEMP1 misfolding and inefficient secretion linked to ML/DHRD, we developed a convenient and sensitive cell-based luminescence assay to monitor secretion versus intracellular accumulation. Fusing EFEMP1 to Gaussia luciferase faithfully recapitulates mutant EFEMP1 secretion defects observed previously using more cumbersome methodology. To understand what governs mutant intracellular retention, we generated a series of R345 mutants. These mutants revealed that aromatic residue substitutions (i.e., Trp, Tyr, and Phe) at position 345 cause significant EFEMP1 secretion deficiencies. These secretion defects appear to be caused, in part, by reduced native disulfide bonding in domain 6 harboring the 345 position. Finally, we demonstrate that mutant EFEMP1 secretion and proper disulfide formation are enhanced by adaptation of the cellular environment by a reduced growth temperature and/or translational attenuation. This study highlights the mechanisms underlying the inefficient secretion of R345W EFEMP1 and demonstrates that alteration of the proteostasis network may provide a strategy to alleviate or delay the onset of this macular dystrophy.

Original languageEnglish (US)
Pages (from-to)4765-4775
Number of pages11
JournalMolecular Biology of the Cell
Volume22
Issue number24
DOIs
StatePublished - Dec 15 2011

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Extracellular Matrix Proteins
Macular Degeneration
Epidermal Growth Factor
Disulfides
Luminescence
fibulin
Luciferases
Homeostasis
Mutation
Temperature
Doyne honeycomb retinal dystrophy
Growth
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration. / Hulleman, John D.; Kaushal, Shalesh; Balch, William E.; Kelly, Jeffery W.

In: Molecular Biology of the Cell, Vol. 22, No. 24, 15.12.2011, p. 4765-4775.

Research output: Contribution to journalArticle

Hulleman, John D. ; Kaushal, Shalesh ; Balch, William E. ; Kelly, Jeffery W. / Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration. In: Molecular Biology of the Cell. 2011 ; Vol. 22, No. 24. pp. 4765-4775.
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