Noncovalent inhibitors reveal BTK gatekeeper and auto-inhibitory residues that control its transforming activity

• DOI: 10.1172/jci.insight.127566
• Publication Date: 2019-06-19T15:01:08Z
• Authors: Shenqiu Wang; Sayan Mondal; Chunying Zhao; Marjan Berishaj; Phani Ghanakota; Connie Lee Batlevi; Ahmet Dogan; Venkatraman E. Se...; Robert Abel; Michael R. Green; Anas Younes; Hans-Guido Wendel
• Keywords: 0301 basic medicine; Binding Sites; Cell Line; 3. Good health; Mice; Structure-Activity Relationship; 03 medical and health sciences; Cell Transformation, Neoplastic; HEK293 Cells; Protein Domains; Mutagenesis; Agammaglobulinaemia Tyrosine Kinase; Animals; Humans; Cysteine; Enzyme Inhibitors

Inhibition of Bruton tyrosine kinase (BTK) is a breakthrough therapy for certain B cell lymphomas and chronic lymphatic leukemia. Covalent BTK inhibitors (e.g., ibrutinib) bind to cysteine C481, mutations this residue confer clinical resistance. This has led the development noncovalent that do not require binding C481. These new compounds are now entering trials. In systematic mutagenesis screen, we identify residues critical activity inhibitors. include gatekeeper (T474) in domain. Strikingly, co-occurrence domain lesions (L512M, E513G, F517L, L547P) cis results 10- 15-fold gain de novo transforming potential vitro vivo. Computational structure analyses reveal how these disrupt an intramolecular mechanism attenuates activation. Our findings anticipate resistance mechanisms class constrain BTK's potential.