Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor

S. Yoshikawa, M. Kukimoto-Niino, L. Parker, N. Handa, T. Terada, T. Fujimoto, Y. Terazawa, M. Wakiyama, M. Sato, S. Sano, T. Kobayashi, T. Tanaka, L. Chen, Z. J. Liu, B. C. Wang, M. Shirouzu, S. Kawa, Kentaro Senba, T. Yamamoto, S. Yokoyama

    Research output: Contribution to journalArticle

    56 Citations (Scopus)

    Abstract

    The epidermal growth factor receptor (EGFR) has an essential role in multiple signaling pathways, including cell proliferation and migration, through extracellular ligand binding and subsequent activation of its intracellular tyrosine kinase (TK) domain. The non-small cell lung cancer (NSCLC)-associated EGFR mutants, L858R and G719S, are constitutively active and oncogenic. They display sensitivity to TK inhibitors, including gefitinib and erlotinib. In contrast, the secondary mutation of the gatekeeper residue, T790M, reportedly confers inhibitor resistance on the oncogenic EGFR mutants. In this study, our biochemical analyses revealed that the introduction of the T790M mutation confers gefitinib resistance on the G719S mutant. The G719S/T790M double mutant has enhanced activity and retains high gefitinib-binding affinity. The T790M mutation increases the ATP affinity of the G719S mutant, explaining the acquired drug resistance of the double mutant. Structural analyses of the G719S/T790M double mutant, as well as the wild type and the G719S and L858R mutants, revealed that the T790M mutation stabilizes the hydrophobic spine of the active EGFR-TK conformation. The Met790 side chain of the G719S/T790M double mutant, in the apo form and gefitinib- and AMPPNP-bound forms, adopts different conformations that explain the accommodation of these ligands. In the L858R mutant structure, the active-site cleft is expanded by the repositioning of Phe723 within the P-loop. Notably, the introduction of the F723A mutation greatly enhanced the gefitinib sensitivity of the wild-type EGFR in vivo, supporting our hypothesis that the expansion of the active-site cleft results in enhanced gefitinib sensitivity. Taken together, our results provide a structural basis for the altered drug sensitivities caused by distinct NSCLC-associated EGFR mutations.

    Original languageEnglish
    Pages (from-to)27-38
    Number of pages12
    JournalOncogene
    Volume32
    Issue number1
    DOIs
    Publication statusPublished - 2013 Jan 3

    Fingerprint

    Epidermal Growth Factor Receptor
    Non-Small Cell Lung Carcinoma
    Mutation
    Pharmaceutical Preparations
    Protein-Tyrosine Kinases
    Catalytic Domain
    Adenylyl Imidodiphosphate
    Ligands
    Drug Resistance
    Cell Movement
    gefitinib
    Spine
    Adenosine Triphosphate
    Cell Proliferation

    Keywords

    • drug resistance
    • EGFR
    • gefitinib
    • NSCLC
    • T790M
    • tyrosine kinase inhibitor

    ASJC Scopus subject areas

    • Molecular Biology
    • Cancer Research
    • Genetics

    Cite this

    Yoshikawa, S., Kukimoto-Niino, M., Parker, L., Handa, N., Terada, T., Fujimoto, T., ... Yokoyama, S. (2013). Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor. Oncogene, 32(1), 27-38. https://doi.org/10.1038/onc.2012.21

    Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor. / Yoshikawa, S.; Kukimoto-Niino, M.; Parker, L.; Handa, N.; Terada, T.; Fujimoto, T.; Terazawa, Y.; Wakiyama, M.; Sato, M.; Sano, S.; Kobayashi, T.; Tanaka, T.; Chen, L.; Liu, Z. J.; Wang, B. C.; Shirouzu, M.; Kawa, S.; Senba, Kentaro; Yamamoto, T.; Yokoyama, S.

    In: Oncogene, Vol. 32, No. 1, 03.01.2013, p. 27-38.

    Research output: Contribution to journalArticle

    Yoshikawa, S, Kukimoto-Niino, M, Parker, L, Handa, N, Terada, T, Fujimoto, T, Terazawa, Y, Wakiyama, M, Sato, M, Sano, S, Kobayashi, T, Tanaka, T, Chen, L, Liu, ZJ, Wang, BC, Shirouzu, M, Kawa, S, Senba, K, Yamamoto, T & Yokoyama, S 2013, 'Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor', Oncogene, vol. 32, no. 1, pp. 27-38. https://doi.org/10.1038/onc.2012.21
    Yoshikawa, S. ; Kukimoto-Niino, M. ; Parker, L. ; Handa, N. ; Terada, T. ; Fujimoto, T. ; Terazawa, Y. ; Wakiyama, M. ; Sato, M. ; Sano, S. ; Kobayashi, T. ; Tanaka, T. ; Chen, L. ; Liu, Z. J. ; Wang, B. C. ; Shirouzu, M. ; Kawa, S. ; Senba, Kentaro ; Yamamoto, T. ; Yokoyama, S. / Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor. In: Oncogene. 2013 ; Vol. 32, No. 1. pp. 27-38.
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    AU - Terada, T.

    AU - Fujimoto, T.

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    AU - Sato, M.

    AU - Sano, S.

    AU - Kobayashi, T.

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    AU - Liu, Z. J.

    AU - Wang, B. C.

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