EDEM1's mannosidase-like domain binds ERAD client proteins in a redox-sensitive manner and possesses catalytic activity

• Published in: The Journal of biological chemistry 2018-09, Vol.293 (36), p.13932-13945
• Main Authors: Lamriben, Lydia, Oster, Michela E., Tamura, Taku, Tian, Weihua, Yang, Zhang, Clausen, Henrik, Hebert, Daniel N.
• Format: Article
• Language: English
• Subjects: alpha-Mannosidase; Animals; Calcium-Binding Proteins; Catalysis; Endoplasmic Reticulum-Associated Degradation; Glycoproteins; HSP40 Heat-Shock Proteins - metabolism; Humans; Mannosidases; Membrane Proteins - metabolism; Molecular Chaperones - metabolism; Oxidation-Reduction; Protein Binding; Protein Domains; Protein Synthesis and Degradation
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA118.004183

Endoplasmic reticulum (ER) degradation-enhancing α-mannosidase–like 1 protein (EDEM1) is a protein quality control factor that was initially proposed to recognize N-linked glycans on misfolded proteins through its mannosidase-like domain (MLD). However, recent studies have demonstrated that EDEM1 binds to some misfolded proteins in a glycan-independent manner, suggesting a more complex binding landscape for EDEM1. In this study, we have identified a thiol-dependent substrate interaction between EDEM1 and the α1-antitrypsin ER-associated protein degradation (ERAD) clients Z and NHK, specifically through the single Cys residue on Z/NHK (Cys256), required for binding under stringent detergent conditions. In addition to the thiol-dependent interaction, the presence of weaker protein-protein interactions was confirmed, suggestive of bipartite client-binding properties. About four reactive thiols on EDEM1 were identified and were not directly responsible for the observed redox-sensitive binding by EDEM1. Moreover, a protein construct comprising the EDEM1 MLD had thiol-dependent binding properties along with its active glycan-trimming activities. Lastly, we identified an additional intrinsically disordered region (IDR) located at the C terminus of EDEM1 in addition to its previously identified N-terminal IDR. We also determined that both IDRs are required for binding to the ERAD component ERdj5 as an interaction with ERdj5 was not observed with the MLD alone. Together, our findings indicate that EDEM1 employs different binding modalities to interact with ERAD clients and ER quality control (ERQC) machinery partners and that some of these properties are shared with its homologues EDEM2 and EDEM3.