A cytosolic reductase pathway is required for efficient N-glycosylation of an STT3B-dependent acceptor site

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dc.contributor.authorvan Lith, Marcel
dc.contributor.authorPringle, Marie Anne
dc.contributor.authorFleming, Bethany
dc.contributor.authorGaeta, Giorgia
dc.contributor.authorIm, Jisu
dc.contributor.authorGilmore, Reid
dc.contributor.authorBulleid, Neil J
dc.contributor.departmentBiochemistry and Molecular Biotechnologyen_US
dc.date.accessioned2023-03-20T13:13:55Z
dc.date.available2023-03-20T13:13:55Z
dc.date.issued2021-11-25
dc.description.abstractN-linked glycosylation of proteins entering the secretory pathway is an essential modification required for protein stability and function. Previously, it has been shown that there is a temporal relationship between protein folding and glycosylation, which influences the occupancy of specific glycosylation sites. Here, we used an in vitro translation system that reproduces the initial stages of secretory protein translocation, folding and glycosylation under defined redox conditions. We found that the efficiency of glycosylation of hemopexin was dependent upon a robust NADPH-dependent cytosolic reductive pathway, which could be mimicked by the addition of a membrane-impermeable reducing agent. We identified a hypoglycosylated acceptor site that is adjacent to a cysteine involved in a short-range disulfide. We show that efficient glycosylation at this site is influenced by the cytosolic reductive pathway acting on both STT3A- and STT3B-dependent glycosylation. Our results provide further insight into the important role of the endoplasmic reticulum redox conditions in glycosylation site occupancy and demonstrate a link between redox conditions in the cytosol and glycosylation efficiency.en_US
dc.identifier.citationvan Lith M, Pringle MA, Fleming B, Gaeta G, Im J, Gilmore R, Bulleid NJ. A cytosolic reductase pathway is required for efficient N-glycosylation of an STT3B-dependent acceptor site. J Cell Sci. 2021 Nov 15;134(22):jcs259340. doi: 10.1242/jcs.259340. Epub 2021 Nov 25. PMID: 34734627; PMCID: PMC8645230.en_US
dc.identifier.doi10.1242/jcs.259340en_US
dc.identifier.eissn1477-9137
dc.identifier.journalJournal of cell science
dc.identifier.pmid34734627
dc.identifier.urihttps://hdl.handle.net/20.500.14038/51854
dc.language.isoenen_US
dc.relation.ispartofJournal of Cell Scienceen_US
dc.relation.urlhttps://doi.org/10.1242/jcs.259340en_US
dc.rights© 2021. Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.en_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.source.countryUnited Kingdom
dc.source.countryUnited Kingdom
dc.source.countryEngland
dc.source.issue22
dc.source.journaltitleJournal of cell science
dc.source.volume134
dc.subjectDisulfide formationen_US
dc.subjectGlycosylationen_US
dc.subjectMagT1en_US
dc.subjectOligosaccharyl transferaseen_US
dc.subjectSTT3Aen_US
dc.subjectSTT3Ben_US
dc.subjectTUSC3en_US
dc.titleA cytosolic reductase pathway is required for efficient N-glycosylation of an STT3B-dependent acceptor siteen_US
dc.typeJournal Articleen_US
dspace.entity.typePublication
refterms.dateFOA2023-03-20T13:13:56Z
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