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Activation of tyrosine kinases by mutation of the gatekeeper threonine

Mohammad Azam1, Markus A Seeliger, Nathanael S Gray

  • 1Karp research building, 7th floor, Division of Pediatric Hematology/Oncology, Children's Hospital of Boston, Massachusetts 02115, USA.

Nature Structural & Molecular Biology
|September 17, 2008
PubMed

Insights

Gatekeeper threonine mutations in tyrosine kinases activate these enzymes, promoting cancer. A novel inhibitor targeting the disrupted hydrophobic spine effectively blocks resistant BCR-ABL mutations.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • Small-molecule inhibitors targeting protein kinases are crucial cancer therapeutics.
  • Resistance to these inhibitors often arises from mutations in the kinase active site, particularly at the gatekeeper residue.

Purpose of the Study:

  • To investigate the role of gatekeeper threonine mutations in tyrosine kinase activation and oncogenesis.
  • To elucidate the structural mechanisms underlying gatekeeper-mediated kinase activation.
  • To develop novel inhibitors targeting resistant kinase mutants.

Main Methods:

  • Investigated gatekeeper mutations in cellular forms of c-ABL, c-SRC, PDGFRA, PDFGRB, and EGFR.
  • Utilized structural analysis to characterize the impact of mutations on kinase conformation.
  • Developed and tested a novel small-molecule inhibitor (compound 14) against resistant mutants.

Main Results:

  • Gatekeeper mutations activate c-ABL, c-SRC, PDGFRA, PDFGRB, and EGFR, promoting malignant transformation of BaF3 cells.
  • Structural analysis revealed stabilization of the active kinase conformation's hydrophobic spine by gatekeeper substitutions.
  • Substitution of glycine for spine residues disrupted hydrophobic interactions and inactivated the kinase.
  • Compound 14 demonstrated efficacy in inhibiting the gatekeeper mutation of BCR-ABL-T315I.

Conclusions:

  • Gatekeeper threonine mutation is a common activation mechanism for tyrosine kinases.
  • The hydrophobic spine is critical for maintaining active kinase conformation.
  • Structural insights from this study can guide the development of next-generation kinase inhibitors effective against resistant mutations.

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