N-terminus-independent activation of c-Src via binding to a tetraspan(in) TM4SF5 in hepatocellular carcinoma is abolished by the TM4SF5 C-terminal peptide application

Haeng Eun Song, Yoonji Lee, Eunmi Kim, Chang Yun Cho, Oisun Jung, Doohyung Lee, Eun Goo Lee, Seo Hee Nam, Minkyung Kang, Stephani Joy Y. Macalino, Ji Eon Kim, Jae Woo Jung, Sung Won Kwon, Sun Choi, Jung Weon Lee

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Active c-Src non-receptor tyrosine kinase localizes to the plasma membrane via N-terminal lipid modification. Membranous c-Src causes cancer initiation and progression. Even though transmembrane 4 L six family member 5 (TM4SF5), a tetraspan(in), can be involved in this mechanism, the molecular and structural influence of TM4SF5 on c-Src remains unknown. Methods: Here, we investigated molecular and structural details by which TM4SF5 regulated c-Src devoid of its N-terminus and how cell-penetrating peptides were able to interrupt c-Src activation via interference of c-Src-TM4SF5 interaction in hepatocellular carcinoma models. Results: The TM4SF5 C-terminus efficiently bound the c-Src SH1 kinase domain, efficiently to the inactively-closed form. The complex involved protein tyrosine phosphatase 1B able to dephosphorylate Tyr530. The c-Src SH1 domain alone, even in a closed form, bound TM4SF5 to cause c-Src Tyr419 and FAK Y861 phosphorylation. Homology modeling and molecular dynamics simulation studies predicted the directly interfacing residues, which were further validated by mutational studies. Cell penetration of TM4SF5 C-terminal peptides blocked the interaction of TM4SF5 with c-Src and prevented c-Src-dependent tumor initiation and progression in vivo. Conclusions: Collectively, these data demonstrate that binding of the TM4SF5 C-terminus to the kinase domain of inactive c-Src leads to its activation. Because this binding can be abolished by cell-penetrating peptides containing the TM4SF5 C-terminus, targeting this direct interaction may be an effective strategy for developing therapeutics that block the development and progression of hepatocellular carcinoma.

Original languageEnglish
Pages (from-to)8092-8111
Number of pages20
Issue number16
StatePublished - 2021

Bibliographical note

Funding Information:
We appreciate Professor Marc Diederich for English proofreading. We also thank the Korea Institute of Science and Technology Information (KISTI) Supercomputing Center for providing computing resources. This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2020R1I1A1A01070020 to EMK, NRF-2018 M3A9C8020027, NRF-2020R1A2C3008993, and NRF-2021M3A9D3024752 to JWL), and the Mid-career Researcher Program (NRF-2020R1A2C2101636), Medical Research Center (MRC) grant (No. 2018R1A5A2025286), and Bio & Medical Technology Development Program (NRF-2019M3E5D4065251) funded by the Ministry of Science and ICT (MSIT) and the Ministry of Health and Welfare (MOHW) through the National Research Foundation of Korea (NRF) to SC.

Publisher Copyright:
© 2021 Ivyspring International Publisher. All rights reserved.


  • Metastasis
  • Protein-protein interaction
  • TM4SF5
  • c-Src


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