C-mannosylation supports folding and enhances stability of thrombospondin repeats.
Authors
Pujols, Jordi
Tiemann, Birgit
Publication Date
2019-12-23Journal Title
Elife
ISSN
2050-084X
Publisher
eLife Sciences Publications, Ltd
Language
en
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Shcherbakova, A., Preller, M., Taft, M. H., Pujols, J., Ventura, S., Tiemann, B., Buettner, F. F., & et al. (2019). C-mannosylation supports folding and enhances stability of thrombospondin repeats.. Elife https://doi.org/10.7554/eLife.52978
Abstract
Previous studies demonstrated importance of C-mannosylation for efficient protein secretion. To study its impact on protein folding and stability, we analyzed both C-mannosylated and non-C-mannosylated thrombospondin type 1 repeats (TSRs) of netrin receptor UNC-5. In absence of C-mannosylation, UNC-5 TSRs could only be obtained at low temperature and a significant proportion displayed incorrect intermolecular disulfide bridging, which was hardly observed when C-mannosylated. Glycosylated TSRs exhibited higher resistance to thermal and reductive denaturation processes, and the presence of C-mannoses promoted the oxidative folding of a reduced and denatured TSR in vitro. Molecular dynamics simulations supported the experimental studies and showed that C-mannoses can be involved in intramolecular hydrogen bonding and limit the flexibility of the TSR tryptophan-arginine ladder. We propose that in the endoplasmic reticulum folding process, C-mannoses orient the underlying tryptophan residues and facilitate the formation of the tryptophan-arginine ladder, thereby influencing the positioning of cysteines and disulfide bridging.
Keywords
Research Article, Biochemistry and Chemical Biology, Cell Biology, glycosylation, thrombospondin type 1 repeats, C-mannosylation, protein folding, protein stability, tryptophan-arginine ladder, C. elegans, D. melanogaster
Sponsorship
Deutsche Forschungsgemeinschaft (FOR2509 BA 4091/6-1)
Deutsche Forschungsgemeinschaft (BA 4091/5-1)
Deutsche Forschungsgemeinschaft (BU 2920/2-1)
Identifiers
52978
External DOI: https://doi.org/10.7554/eLife.52978
This record's URL: https://www.repository.cam.ac.uk/handle/1810/300786
Rights
Attribution 4.0 International (CC BY 4.0)
Licence URL: https://creativecommons.org/licenses/by/4.0/
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