2. Academic Validation
  3. GW806742X can induce mouse MLKL activation by directly promoting MLKL kinase like domain dimerization

GW806742X can induce mouse MLKL activation by directly promoting MLKL kinase like domain dimerization

  • Exp Cell Res. 2025 May 1;448(1):114539. doi: 10.1016/j.yexcr.2025.114539.
Feiyang Yuan 1,  Yu Zhang 1,  Xinxin Zhu 1,  Hong Hu 1,  Ning Nan 1,  Huayi Wang 2
Affiliations

Affiliations

  • 1 School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China; State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
  • 2 School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China; Beigene, Ltd., Shanghai, 200020, China. Electronic address: huayi.wang@beigene.com.
PMID: 40189182 DOI: 10.1016/j.yexcr.2025.114539
Abstract

The dimerization of the MLKL kinase-like domain (KLD) is a crucial step for MLKL activation in Necroptosis. In 2014, it was discovered that GW806742X can directly bind to the mouse MLKL KLD via surface plasmon resonance (SPR) (Kd = 9.3 μM), inhibiting TNF-induced membrane translocation of MLKL and Necroptosis. Consequently, GW806742X is considered a mouse MLKL inhibitor. In this study, we found that GW806742X blocks TNF-induced RIP1-dependent Necroptosis but promotes Necroptosis triggered by either RIP3 or MLKL self-oligomerization in the FKBPv chemical dimerizer system. In addition, higher doses of GW806742X can directly induce MLKL-dependent Necroptosis and promote MLKL oligomerization, as detected by non-reducing Western blot. Through chemical cross-linking assays, we observed that GW806742X induces dimerization of recombinant mouse MLKL KLD proteins. The dimerization of the MLKL KLD is a direct consequence of RIP3 phosphorylation, a crucial step in RIP3-induced MLKL activation and Necroptosis. Therefore, GW806742X exerts a dual effect on necroptosis: it inhibits Necroptosis, likely by interfering with RIP1 function, while promoting Necroptosis by facilitating MLKL activation.

Keywords

GW806742X; Kinase-like domain; MLKL; Necroptosis.

Figures
Products