NMR spectroscopic characterization of the C‐mannose conformation in a thrombospondin repeat using a selective labeling approach
- Despite the great interest in glycoproteins, structural information reporting on conformation and dynamics of the sugar moieties are limited. We present a new biochemical method to express proteins with glycans that are selectively labeled with NMR‐active nuclei. We report on the incorporation of 13C‐labeled mannose in the C‐mannosylated UNC‐5 thrombospondin repeat. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of UNC‐5 is characterized by interconversion between the canonical 1C4 state and the B03 / 1S3 state. This flexibility may be essential for protein folding and stabilization. We foresee that this versatile tool to produce proteins with selectively labeled C‐mannose can be applied and adjusted to other systems and modifications and potentially paves a way to advance glycoprotein research by unravelling the dynamical and conformational properties of glycan structures and their interactions.
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| Author: | Hendrik R. A. JonkerORCiD, Krishna SaxenaORCiDGND, Aleksandra Shcherbakova, Birgit Tiemann, Hans Bakker, Harald SchwalbeORCiDGND |
|---|---|
| URN: | urn:nbn:de:hebis:30:3-570190 |
| DOI: | https://doi.org/10.1002/anie.202009489 |
| ISSN: | 1521-3773 |
| ISSN: | 1433-7851 |
| Parent Title (German): | Angewandte Chemie. International edition |
| Publisher: | Wiley-VCH |
| Place of publication: | Weinheim |
| Document Type: | Article |
| Language: | English |
| Date of Publication (online): | 2020/08/03 |
| Date of first Publication: | 2020/08/03 |
| Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
| Release Date: | 2020/12/04 |
| Tag: | C-mannosylation; NMR spectroscopy; conformation analysis; glycoproteins; selective isotopic labeling |
| Volume: | 59 |
| Issue: | 46 |
| Page Number: | 7 |
| First Page: | 20659 |
| Last Page: | 20665 |
| HeBIS-PPN: | 476110718 |
| Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
| Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
| 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie | |
| Sammlungen: | Universitätspublikationen |
| Licence (German): |  Creative Commons - Namensnennung 4.0 |
