A role for N-glycanase in the cytosolic turnover of glycoproteins

• Published in: The EMBO journal Vol. 22; no. 5; pp. 1036 - 1046
• Main Authors: Hirsch, Christian, Blom, Daniël, Ploegh, Hidde L.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 03-03-2003; Blackwell Publishing Ltd; Oxford University Press
• Subjects: Amidohydrolases - genetics; Amidohydrolases - metabolism; Animals; Binding Sites; Cell Line; Cysteine - metabolism; Cysteine Endopeptidases - chemistry; Cysteine Endopeptidases - metabolism; Cytoplasm - enzymology; Endoplasmic Reticulum - metabolism; ER quality control; Fungal Proteins - genetics; Fungal Proteins - metabolism; glycoprotein; Glycoproteins; Glycoproteins - metabolism; Glycosylation; Histocompatibility Antigens Class I - chemistry; Histocompatibility Antigens Class I - metabolism; Humans; Mammals; Models, Biological; Models, Molecular; Molecular Sequence Data; Multienzyme Complexes - chemistry; Multienzyme Complexes - metabolism; N-glycanase; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; proteasome; Proteasome Endopeptidase Complex; Protein Structure, Tertiary; Protein Transport - physiology; Receptors, Antigen, T-Cell, alpha-beta - metabolism; siRNA; Yeasts
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1093/emboj/cdg107

Successful maturation determines the intracellular fate of secretory and membrane proteins in the endoplasmic reticulum (ER). Failure of proteins to fold or assemble properly can lead to their retention in the ER and redirects them to the cytosol for degradation by the proteasome. Proteasome inhibitors can yield deglycosylated cytoplasmic intermediates that are the result of an N‐glycanase activity, believed to act prior to destruction of these substrates by the proteasome. A gene encoding a yeast peptide:N‐glycanase, PNG1, has been cloned, but this N‐glycanase and its mammalian homolog were reported to be incapable of deglycosylating full‐length glycoproteins. We show that both the yeast PNG1 enzyme and its mammalian homolog display N‐glycanase activity towards intact glycoproteins. As substrates, cytosolic PNGase activity prefers proteins containing high‐mannose over those bearing complex type oligosaccharides. Importantly, PNG1 discriminates between non‐native and folded glycoproteins, consistent with a role for N‐glycanase in cytoplasmic turnover of glycoproteins.