Abstract
NOTCH1 is a transmembrane receptor in metazoans that is linked to a variety of disorders. The receptor contains an extracellular domain (ECD) with 36 tandem epidermal growth factor-like (EGF) repeats. The ECD is responsible for intercellular signaling via protein–ligand interactions with neighboring cells. Each EGF repeat consists of approximately 40 amino acids and 3 conserved disulfide bonds. The Abruptex region (EGF24-29) is critical for NOTCH1 signaling and is known for its missense mutations. Certain EGF repeats are modified with the addition of O-linked glycans and many have calcium binding sites, which give each EGF repeat a unique function. It has been shown that the loss of the O-fucose site of EGF27 alters NOTCH1 activity. To investigate the role of glycosylation in the NOTCH1 signaling pathway, nuclear magnetic resonance spectroscopy has been employed to study the structures of EGF27 and its glycoforms. Here, we report the backbone and sidechain 1H, 15N, and 13C-resonance assignments of the unmodified EGF27 protein and the predicted secondary structure derived from the assigned chemical shifts.
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Data availability
The raw NMR data and assignments are available through the Biological Magnetic Resonance Data Bank (http://www.bmrb.wisc.edu/), Accession Number 50703.
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Acknowledgements
This work was supported by funding from the National Institute of General Medical Sciences (GM061126, Robert S. Haltiwanger) and the National Science Foundation (CHE-1413576 and BIO-2019046, Megan A. Macnaughtan). Justin A. Grennell thanks the Louisiana State University Department of Chemistry for providing support through teaching assistantships. The authors thank Dr. Woonghee Lee for software assistance with NMRFAM-SPARKY.
Funding
This work was supported by funding from the National Institute of General Medical Sciences (GM061126, RSH) and the National Science Foundation (CHE-1413576, MAM). JAG thanks the Louisiana State University Department of Chemistry for providing support through teaching assistantships.
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JAG, KDJ, and KBL prepared materials and contributed to the experimental design with MAM and RSH. JAG and JG collected and processed the NMR data. JAG and MAM analyzed and interpreted the NMR data. The first draft of the manuscript was written by JAG and edited by MAM and RSH. All authors read and approved the final manuscript.
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Grennell, J.A., Jenkins, K.D., Luther, K.B. et al. 1H, 15N, 13C backbone and sidechain resonance assignments and secondary structure of mouse NOTCH1 EGF27. Biomol NMR Assign 17, 27–35 (2023). https://doi.org/10.1007/s12104-022-10116-0
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DOI: https://doi.org/10.1007/s12104-022-10116-0