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Cited by 18Biotin and Related StudiesHER2/EGFR in Cancer ResearchProtein Degradation and Inhibitors

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Proximity extracellular protein-protein interaction analysis of EGFR using AirID-conjugated fragment of antigen binding

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Yamada, Kohdai, Shioya, Ryouhei, Nishino, Kohei, Furihata, Hirotake, Hijikata, Atsushi, Kaneko, Mika K., Kato, Yukinari, Shirai, Tsuyoshi, Kosako, Hidetaka, Sawasaki, Tatsuya·2023·Nature Communications
DOI 10.1038/s41467-023-43931-7PMC PMC10721602

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Abstract

Receptor proteins, such as epidermal growth factor receptor (EGFR), interact with other proteins in the extracellular region of the cell membrane to drive intracellular signalling. Therefore, analysis of extracellular protein-protein interactions (exPPIs) is important for understanding the biological function of receptor proteins. Here, we present an approach using a proximity biotinylation enzyme (AirID) fusion fragment of antigen binding (FabID) to analyse the proximity exPPIs of EGFR. AirID was C-terminally fused to the Fab fragment against EGFR (EGFR-FabID), which could then biotinylate the extracellular region of EGFR in several cell lines. Liquid Chromatography-Mass Spectrometry (LC-MS/MS) analysis indicated that many known EGFR interactors were identified as proximity exPPIs, along with many unknown candidate interactors, using EGFR-FabID. Interestingly, these proximity exPPIs were influenced by treatment with EGF ligand and its specific kinase inhibitor, gefitinib. These results indicate that FabID provides accurate proximity exPPI analysis of target receptor proteins on cell membranes with ligand and drug responses.

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Metadata

Authors
Yamada, K., Shioya, R., Nishino, K., Furihata, H., Hijikata, A., Kaneko, M. K., Kato, Y., Shirai, T., Kosako, H., & Sawasaki, T.
Affiliations
  • Ehime University (JP)
  • Institute of Advanced Medical Sciences, Tokushima University (JP)
  • Tokyo University of Pharmacy and Life Sciences (JP)
  • Tohoku University (JP)
  • Nagahama Institute of Bio-Science and Technology (JP)
Corresponding author
  • Kosako, Hidetaka
  • Sawasaki, Tatsuya
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2023-12-14
Volume
14
Issue
1
Pages
8301-8301
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-023-43931-7
PMC
PMC10721602
OpenAlex
W4389738380
Cited by
18
References
2
Citations by year
2026: 9 · 2025: 6 · 2024: 3
FWCI
2.60
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Institutional Repositories DataBase (IRDB) (acceptedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Japan Agency for Medical Research and Development (JP21am0101077, 22ama121010j0001, 23ama121010j0002, JP22ama121008)
  • MEXT | Japan Society for the Promotion of Science (19H03218, JP21K19230)
  • Takeda Science Foundation
  • Joint Usage and Joint Research Programs of the Institute of Advanced Medical Sciences, Tokushima University
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
  • Health Sciences › Medicine › Oncology › HER2/EGFR in Cancer Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
Keywords
epidermal growth factor receptor, proximity biotinylation, receptor signaling, EGF ligands, gefitinib, kinase inhibitors, protein-protein interaction network
MeSH terms
Chromatography, Liquid, Epidermal Growth Factor, Ligands, Phosphorylation, Tandem Mass Spectrometry, ErbB Receptors

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