EDEM3 Domains Cooperate to Perform Its Overall Cell Functioning
| Autor: | Petruta R. Alexandru, Gabriela Chiritoiu, Cristian V.A. Munteanu, Marius Surleac, Georgiana Manica, Simona Ghenea, Andrei-Jose Petrescu, Cristian M. Butnaru, Stefana M. Petrescu, Eliza C. Martin |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Mannosidase Protein Folding Mutant Mannose Endoplasmic Reticulum lcsh:Chemistry chemistry.chemical_compound 0302 clinical medicine Catalytic Domain Protein Interaction Maps lcsh:QH301-705.5 Spectroscopy mass spectrometry chemistry.chemical_classification Chemistry Monophenol Monooxygenase Man1B1 General Medicine Endoplasmic Reticulum-Associated Degradation Computer Science Applications Cell biology ER mannosidases Protein folding Sequence analysis intrinsically disordered domain Protein degradation Endoplasmic-reticulum-associated protein degradation tyrosinase NHK Article Catalysis Inorganic Chemistry 03 medical and health sciences Protein Domains alpha-Mannosidase Mannosidases Humans Physical and Theoretical Chemistry Molecular Biology protease-associated domain Organic Chemistry Calcium-Binding Proteins ERAD 030104 developmental biology HEK293 Cells lcsh:Biology (General) lcsh:QD1-999 alpha 1-Antitrypsin Mutation EDEM3 Glycoprotein 030217 neurology & neurosurgery HeLa Cells |
| Zdroj: | International Journal of Molecular Sciences, Vol 22, Iss 2172, p 2172 (2021) International Journal of Molecular Sciences Volume 22 Issue 4 |
| ISSN: | 1422-0067 |
| Popis: | EDEM3 recognizes and directs misfolded proteins to the ER-associated protein degradation (ERAD) process. EDEM3 was predicted to act as lectin or as a mannosidase because of its homology with the GH47 catalytic domain of the Man1B1, but the contribution of the other regions remained unresolved. Here, we dissect the molecular determinants governing EDEM3 function and its cellular interactions. LC/MS analysis indicates very few stable ER interactors, suggesting EDEM3 availability for transient substrate interactions. Sequence analysis reveals that EDEM3 consists of four consecutive modules defined as GH47, intermediate (IMD), protease-associated (PA), and intrinsically disordered (IDD) domain. Using an EDEM3 knock-out cell line, we expressed EDEM3 and domain deletion mutants to address EDEM3 function. We find that the mannosidase domain provides substrate binding even in the absence of mannose trimming and requires the IMD domain for folding. The PA and IDD domains deletions do not impair the trimming, but specifically modulate the turnover of two misfolded proteins, NHK and the soluble tyrosinase mutant. Hence, we demonstrate that EDEM3 provides a unique ERAD timing to misfolded glycoproteins, not only by its mannose trimming activity, but also by the positive and negative feedback modulated by the protease-associated and intrinsically disordered domain, respectively. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|