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PMID: 9177202 Published · ppublish English Journal Article

Site-specific de-N-glycosylation of diglycosylated ovalbumin in hen oviduct by endogenous peptide: N-glycanase as a quality control system for newly synthesized proteins.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 94 ·No. 12 ·1997-06-10 ·Pages 6244-9

Suzuki T, Kitajima K, Emori Y, Inoue Y, Inoue S

Abstract

Hen ovalbumin (OVA) is known to exist as a singly N-glycosylated form with a glycan chain on Asn-292 in egg white. Previous studies showed that di-N-glycosylated form of OVA [Di-OVA; CHO-Asn-292/CHO-Asn-311 (CHO, N-glycan chain)], which has two N-glycan chains on Asn-292 and Asn-311, was expressed only transiently in hen oviduct. Di-OVA was not found in egg white, suggesting that this form cannot be secreted normally and may possibly be converted to mono-N-glycosylated OVA (CHO-Asn-292/Asp-311) by the action of peptide:N-glycanase (PNGase) during synthesis and secretion. In this study, we have identified the putative PNGase activity in the homogenate of hen oviduct, purified 1,000-fold, and designated as PNGase HO. We examined the reactivity of Di-OVA to PNGase HO and found that this enzyme site-specifically cleaved off the glycan chain at Asn-311 to convert Di-OVA into the mono-N-glycosylated form (CHO-Asn-292/Asp-311). In contrast, this enzyme was found not to act on the mono-N-glycosylated OVA (CHO-Asn-292/Asn-311) found in egg white when it was tested as a substrate. The present findings support our view that de-N-glycosylation catalyzed by PNGase may be involved in quality control of newly synthesized proteins by converting its diglycosylated form into the mono-N-glycosylated form that can be secreted. However, the alternative possibility that de-N-glycosylation may trigger cytosolic degradation of the aberrantly glycosylated glycoprotein cannot be ruled out.

MeSH Terms
Amidohydrolases/isolation & purification,metabolism Animals Asparagine Chickens Chromatography, Gel Chromatography, Ion Exchange Endopeptidases/metabolism Female Glycopeptides/chemistry,isolation & purification Glycoproteins/biosynthesis Glycoside Hydrolases/metabolism Glycosylation Ovalbumin/biosynthesis,chemistry,metabolism Oviducts/metabolism Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Quality Control Substrate Specificity
Chemicals
Glycopeptides Glycoproteins Asparagine Ovalbumin Glycoside Hydrolases Endopeptidases Amidohydrolases Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Suzuki T
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo 113, Japan.
Kitajima K
Emori Y
Inoue Y
Inoue S
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.94.12.6244
Published
1997-06-10
Pages
6244-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC21034
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.94.12.6244 PMC Full Text View journal details
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