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:	Marioara B Marin, Simona Ghenea, Laurentiu N Spiridon, Gabriela N Chiritoiu, Andrei-Jose Petrescu, Stefana-Maria Petrescu
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/102caa9e862244ed938780359f9a13ac
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spelling	oai:doaj.org-article:102caa9e862244ed938780359f9a13ac2021-11-18T07:09:16ZTyrosinase degradation is prevented when EDEM1 lacks the intrinsically disordered region.1932-620310.1371/journal.pone.0042998https://doaj.org/article/102caa9e862244ed938780359f9a13ac2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22905195/?tool=EBIhttps://doaj.org/toc/1932-6203EDEM1 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.Marioara B MarinSimona GheneaLaurentiu N SpiridonGabriela N ChiritoiuGabriela N ChiritoiuAndrei-Jose PetrescuStefana-Maria PetrescuPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 8, p e42998 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Marioara B Marin Simona Ghenea Laurentiu N Spiridon Gabriela N Chiritoiu Gabriela N Chiritoiu Andrei-Jose Petrescu Stefana-Maria Petrescu Tyrosinase degradation is prevented when EDEM1 lacks the intrinsically disordered region.
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.
format	article
author	Marioara B Marin Simona Ghenea Laurentiu N Spiridon Gabriela N Chiritoiu Gabriela N Chiritoiu Andrei-Jose Petrescu Stefana-Maria Petrescu
author_facet	Marioara B Marin Simona Ghenea Laurentiu N Spiridon Gabriela N Chiritoiu Gabriela N Chiritoiu Andrei-Jose Petrescu Stefana-Maria Petrescu
author_sort	Marioara B Marin
title	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_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_sort	tyrosinase degradation is prevented when edem1 lacks the intrinsically disordered region.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/102caa9e862244ed938780359f9a13ac
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_version_	1718423835467841536

Tyrosinase degradation is prevented when EDEM1 lacks the intrinsically disordered region.

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