Lathyrol Targets G6PD to Disrupt Redox Homeostasis in Non-Small Cell Lung Cancer.

Lathyrol, a bioactive natural compound derived from plants of the Euphorbiaceae family, exhibits antitumor activity, and its molecular targets and underlying mechanisms remain incompletely understood. In this study, thermal proteome profiling (TPP) was applied to systematically identify lathyrol-binding proteins in non-small cell lung cancer (NSCLC) cells. TPP analysis identified glucose-6-phosphate dehydrogenase (G6PD) as a candidate target of lathyrol. A cellular thermal shift assay (CETSA) confirmed increased thermal stability of G6PD upon treatment. Molecular docking indicated a potential interaction between lathyrol and G6PD. The peptide-centric local stability assay (PELSA) revealed localized conformational changes in the C-terminal region of G6PD consistent with the predicted interaction interface. Enzymatic assays showed reduced G6PD activity accompanied by decreased intracellular NADPH levels. Quantitative proteomics indicated alterations in the pathways associated with glucose metabolism and redox regulation. These findings identify G6PD as a functional target of lathyrol in NSCLC cells and link its inhibition to disruption of cellular redox balance and metabolic homeostasis.