C-mannosylation supports folding and enhances stability of thrombospondin repeats.
| Autor: | Shcherbakova A; Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany., Preller M; Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany., Taft MH; Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany., Pujols J; Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, Spain., Ventura S; Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, Spain., Tiemann B; Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany., Buettner FF; Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany., Bakker H; Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany. |
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| Jazyk: | angličtina |
| Zdroj: | ELife [Elife] 2019 Dec 23; Vol. 8. Date of Electronic Publication: 2019 Dec 23. |
| DOI: | 10.7554/eLife.52978 |
| Abstrakt: | 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. Competing Interests: AS, MP, MT, JP, SV, BT, FB, HB No competing interests declared (© 2019, Shcherbakova et al.) |
| Databáze: | MEDLINE |
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