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Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region

EDEM1 is a mannosidase-like protein that recruits misfolded glycoproteins from the calnexin/calreticulin folding cycle to downstream endoplasmic reticulum associated degradation (ERAD) pathway. Here, we investigate the role of EDEM1 in the processing of tyrosinase, a tumour antigen overexpressed in...

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Autores principales: Marin, Marioara B., Ghenea, Simona, Spiridon, Laurentiu N., Chiritoiu, Gabriela N., Petrescu, Andrei-Jose, Petrescu, Stefana-Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3414498/
https://www.ncbi.nlm.nih.gov/pubmed/22905195
http://dx.doi.org/10.1371/journal.pone.0042998
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author Marin, Marioara B.
Ghenea, Simona
Spiridon, Laurentiu N.
Chiritoiu, Gabriela N.
Petrescu, Andrei-Jose
Petrescu, Stefana-Maria
author_facet Marin, Marioara B.
Ghenea, Simona
Spiridon, Laurentiu N.
Chiritoiu, Gabriela N.
Petrescu, Andrei-Jose
Petrescu, Stefana-Maria
author_sort Marin, Marioara B.
collection PubMed
description EDEM1 is a mannosidase-like protein that recruits misfolded glycoproteins from the calnexin/calreticulin folding cycle to downstream endoplasmic reticulum associated degradation (ERAD) pathway. Here, we investigate the role of EDEM1 in the processing of tyrosinase, a tumour antigen overexpressed in melanoma cells. First, we analyzed and modeled EDEM1 major domains. The homology model raised on the crystal structures of human and Saccharomyces cerevisiae ER class I α1,2-mannosidases reveals that the major mannosidase domain located between aminoacids 121–598 fits with high accuracy. We have further identified an N-terminal region located between aminoacids 40–119, predicted to be intrinsically disordered (ID) and susceptible to adopt multiple conformations, hence facilitating protein-protein interactions. To investigate these two domains we have constructed an EDEM1 deletion mutant lacking the ID region and a triple mutant disrupting the glycan-binding domain and analyzed their association with tyrosinase. Tyrosinase is a glycoprotein partly degraded endogenously by ERAD and the ubiquitin proteasomal system. We found that the degradation of wild type and misfolded tyrosinase was enhanced when EDEM1 was overexpressed. Glycosylated and non-glycosylated mutants co-immunoprecipitated with EDEM1 even in the absence of its intact mannosidase-like domain, but not when the ID region was deleted. In contrast, calnexin and SEL 1L associated with the deletion mutant. Our data suggest that the ID region identified in the N-terminal end of EDEM1 is involved in the binding of glycosylated and non-glycosylated misfolded proteins. Accelerating tyrosinase degradation by EDEM1 overexpression may lead to an efficient antigen presentation and enhanced elimination of melanoma cells.
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spelling pubmed-34144982012-08-19 Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region Marin, Marioara B. Ghenea, Simona Spiridon, Laurentiu N. Chiritoiu, Gabriela N. Petrescu, Andrei-Jose Petrescu, Stefana-Maria PLoS One Research Article EDEM1 is a mannosidase-like protein that recruits misfolded glycoproteins from the calnexin/calreticulin folding cycle to downstream endoplasmic reticulum associated degradation (ERAD) pathway. Here, we investigate the role of EDEM1 in the processing of tyrosinase, a tumour antigen overexpressed in melanoma cells. First, we analyzed and modeled EDEM1 major domains. The homology model raised on the crystal structures of human and Saccharomyces cerevisiae ER class I α1,2-mannosidases reveals that the major mannosidase domain located between aminoacids 121–598 fits with high accuracy. We have further identified an N-terminal region located between aminoacids 40–119, predicted to be intrinsically disordered (ID) and susceptible to adopt multiple conformations, hence facilitating protein-protein interactions. To investigate these two domains we have constructed an EDEM1 deletion mutant lacking the ID region and a triple mutant disrupting the glycan-binding domain and analyzed their association with tyrosinase. Tyrosinase is a glycoprotein partly degraded endogenously by ERAD and the ubiquitin proteasomal system. We found that the degradation of wild type and misfolded tyrosinase was enhanced when EDEM1 was overexpressed. Glycosylated and non-glycosylated mutants co-immunoprecipitated with EDEM1 even in the absence of its intact mannosidase-like domain, but not when the ID region was deleted. In contrast, calnexin and SEL 1L associated with the deletion mutant. Our data suggest that the ID region identified in the N-terminal end of EDEM1 is involved in the binding of glycosylated and non-glycosylated misfolded proteins. Accelerating tyrosinase degradation by EDEM1 overexpression may lead to an efficient antigen presentation and enhanced elimination of melanoma cells. Public Library of Science 2012-08-08 /pmc/articles/PMC3414498/ /pubmed/22905195 http://dx.doi.org/10.1371/journal.pone.0042998 Text en © 2012 Marin et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Marin, Marioara B.
Ghenea, Simona
Spiridon, Laurentiu N.
Chiritoiu, Gabriela N.
Petrescu, Andrei-Jose
Petrescu, Stefana-Maria
Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region
title Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region
title_full Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region
title_fullStr Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region
title_full_unstemmed Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region
title_short Tyrosinase Degradation Is Prevented when EDEM1 Lacks the Intrinsically Disordered Region
title_sort tyrosinase degradation is prevented when edem1 lacks the intrinsically disordered region
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3414498/
https://www.ncbi.nlm.nih.gov/pubmed/22905195
http://dx.doi.org/10.1371/journal.pone.0042998
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