EDEM accelerates ERAD by preventing aberrant dimer formation of misfolded alpha1-antitrypsin.
Misfolded glycoproteins are degraded by a mechanism known as ERAD (ER-associated degradation) after retrotranslocation out of the endoplasmic reticulum (ER). This mechanism plays an important role in ER quality control. We previously reported that an ER membrane protein, EDEM, accelerates ERAD of a misfolded alpha1-antitrypsin variant, null (Hong Kong) (NHK), suggesting that EDEM may function as an acceptor of terminally misfolded glycoproteins. In this study, we constructed several genetically manipulated cell lines to test this hypothesis. EDEM expression did not alter the secretion rate of properly folded molecules and the forced retention of wild-type alpha1-antitrypsin in the ER did not cause its association with EDEM, suggesting that EDEM may function as a molecular chaperone. To examine this possibility, we analyzed the effect of EDEM over-expression on the structure of NHK, and found that the accumulation of covalent NHK dimers was selectively prevented by the over-expression of EDEM. Co-expression of NHK with two other ER membrane proteins, calnexin and H(+)/K(+)-ATPase (beta subunit), did not inhibit NHK dimer formation or accelerate NHK ERAD. These results indicate that EDEM may maintain the retrotranslocation competence of NHK by inhibiting aggregation so that unstable misfolded proteins can be accommodated by the dislocon for ERAD.
Hosokawa
Nobuko
N
Department of Molecular and Cellular Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8397, Japan. nobuko@frontier.kyoto-u.ac.jp
Wada
Ikuo
I
Natsuka
Yuko
Y
Nagata
Kazuhiro
K
D000251:Adenosine Triphosphatases / Q000378:metabolism; D037281:Calnexin / Q000378:metabolism / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D019281:Dimerization; D004229:Dithiothreitol / Q000378:metabolism / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism* / Q000494:pharmacology*; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D021122:Protein Subunits / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
C486654:EDEM1 protein, human; D006023:Glycoproteins; D008565:Membrane Proteins; D021122:Protein Subunits; D000515:alpha 1-Antitrypsin; D037281:Calnexin; D010088:Oxidoreductases; D000251:Adenosine Triphosphatases; D004229:Dithiothreitol