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

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 homeostasi...

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Autores principales: Hulleman, John D., Kaushal, Shalesh, Balch, William E., Kelly, Jeffery W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3237620/
https://www.ncbi.nlm.nih.gov/pubmed/22031286
http://dx.doi.org/10.1091/mbc.E11-08-0695
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author Hulleman, John D.
Kaushal, Shalesh
Balch, William E.
Kelly, Jeffery W.
author_facet Hulleman, John D.
Kaushal, Shalesh
Balch, William E.
Kelly, Jeffery W.
author_sort Hulleman, John D.
collection PubMed
description 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.
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spelling pubmed-32376202012-03-01 Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration Hulleman, John D. Kaushal, Shalesh Balch, William E. Kelly, Jeffery W. Mol Biol Cell Articles 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. The American Society for Cell Biology 2011-12-15 /pmc/articles/PMC3237620/ /pubmed/22031286 http://dx.doi.org/10.1091/mbc.E11-08-0695 Text en © 2011 Hulleman et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Hulleman, John D.
Kaushal, Shalesh
Balch, William E.
Kelly, Jeffery W.
Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration
title Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration
title_full Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration
title_fullStr Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration
title_full_unstemmed Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration
title_short Compromised mutant EFEMP1 secretion associated with macular dystrophy remedied by proteostasis network alteration
title_sort compromised mutant efemp1 secretion associated with macular dystrophy remedied by proteostasis network alteration
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3237620/
https://www.ncbi.nlm.nih.gov/pubmed/22031286
http://dx.doi.org/10.1091/mbc.E11-08-0695
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