C-mannosylation supports folding and enhances stability of thrombospondin repeats.
eLife Sciences Publications, Ltd
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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
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.
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
Deutsche Forschungsgemeinschaft (FOR2509 BA 4091/6-1)
Deutsche Forschungsgemeinschaft (BA 4091/5-1)
Deutsche Forschungsgemeinschaft (BU 2920/2-1)
External DOI: https://doi.org/10.7554/eLife.52978
This record's URL: https://www.repository.cam.ac.uk/handle/1810/300786
Attribution 4.0 International (CC BY 4.0)
Licence URL: https://creativecommons.org/licenses/by/4.0/