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| DOI | Resolve DOI: https://doi.org/10.1038/s41598-021-83198-w |
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| Author | Search for: Sheff, Joey1; Search for: Wang, Ping1; Search for: Xu, Ping1; Search for: Arbour, Melanie1; Search for: Masson, Luke1; Search for: Van Faassen, Henk1; Search for: Hussack, Greg1; Search for: Kemmerich, Kristin1; Search for: Brunette, Eric1; Search for: Stanimirovic, Danica1; Search for: Hill, Jennifer J.1; Search for: Kelly, John1; Search for: Ni, Feng1 |
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| Affiliation | - National Research Council of Canada. Human Health Therapeutics
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| Format | Text, Article |
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| Subject | biochemistry; drug discovery; structural biology |
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| Abstract | Ligand-activated signaling through the type 1 insulin-like growth factor receptor (IGF1R) is implicated in many physiological processes ranging from normal human growth to cancer proliferation and metastasis. IGF1R has also emerged as a target for receptor-mediated transcytosis, a transport phenomenon that can be exploited to shuttle biotherapeutics across the blood–brain barrier (BBB). We employed differential hydrogen–deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) to characterize the interactions of the IGF1R ectodomain with a recently discovered BBB-crossing single-domain antibody (sdAb), VHH-IR5, in comparison with IGF-1 binding. HDX-MS confirmed that IGF-1 induced global conformational shifts in the L1/FnIII-1/-2 domains and α-CT helix of IGF1R. In contrast, the VHH-IR5 sdAb-mediated changes in conformational dynamics were limited to the α-CT helix and its immediate vicinity (L1 domain). High-resolution NMR spectroscopy titration data and linear peptide scanning demonstrated that VHH-IR5 has high-affinity binding interactions with a peptide sequence around the C-terminal region of the α-CT helix. Taken together, these results define a core linear epitope for VHH-IR5 within the α-CT helix, overlapping the IGF-1 binding site, and suggest a potential role for the α-CT helix in sdAb-mediated transcytosis. |
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| Publication date | 2021-02-19 |
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| Publisher | Nature Research |
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| Licence | |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | e728127d-7bb4-4c3c-9f37-02ade41abddc |
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| Record created | 2021-04-16 |
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| Record modified | 2021-04-20 |
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