EDEM3, a soluble EDEM homolog, enhances glycoprotein endoplasmic reticulum-associated degradation and mannose trimming.
Quality control in the endoplasmic reticulum ensures that only properly folded proteins are retained in the cell through mechanisms that recognize and discard misfolded or unassembled proteins in a process called endoplasmic reticulum-associated degradation (ERAD). We previously cloned EDEM (ER degradation-enhancing alpha-mannosidase-like protein) and showed that it accelerates ERAD of misfolded glycoproteins. We now cloned mouse EDEM3, a soluble homolog of EDEM. EDEM3 consists of 931 amino acids and has all the signature motifs of Class I alpha-mannosidases (glycosyl hydrolase family 47) in its N-terminal domain and a protease-associated motif in its C-terminal region. EDEM3 accelerates glycoprotein ERAD in transfected HEK293 cells, as shown by increased degradation of misfolded alpha1-antitrypsin variant (null (Hong Kong)) and of TCRalpha. Overexpression of EDEM3 also greatly stimulates mannose trimming not only from misfolded alpha1-AT null (Hong Kong) but also from total glycoproteins, in contrast to EDEM, which has no apparent alpha1,2-mannosidase activity. Furthermore, overexpression of the E147Q EDEM3 mutant, which has the mutation in one of the conserved acidic residues essential for enzyme activity of alpha1,2-mannosidases, abolishes the stimulation of mannose trimming and greatly decreases the stimulation of ERAD by EDEM3. These results show that EDEM3 has alpha1,2-mannosidase activity in vivo, suggesting that the mechanism whereby EDEM3 accelerates glycoprotein ERAD is different from that of EDEM.
Hirao
Kazuyoshi
K
Department of Molecular and Cellular Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8397, Japan, CREST, JST, Saitama 332-0012, Japan.
Natsuka
Yuko
Y
Tamura
Taku
T
Wada
Ikuo
I
Morito
Daisuke
D
Natsuka
Shunji
S
Romero
Pedro
P
Sleno
Barry
B
Tremblay
Linda O
LO
Herscovics
Annette
A
Nagata
Kazuhiro
K
Hosokawa
Nobuko
N
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000502:physiology*; D020224:Expressed Sequence Tags; D006023:Glycoproteins / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D017510:Protein Folding; D012995:Solubility; D043323:alpha-Mannosidase / Q000378:metabolism
D002135:Calcium-Binding Proteins; D006023:Glycoproteins; D008565:Membrane Proteins; C510458:EDEM3 protein, mouse; D043323:alpha-Mannosidase; D008358:Mannose