A multi-disciplinary strategy has been developed and applied for identifying Tatridin A target(s) to fully address its mechanism of action, based on drug affinity response target stability and targeted limited proteolysis. Both approaches point to the identification of PGK 1 and 2 (phospoglicerate kinase 1 and 2) as major Tatridin A targets in THP-1 cell lysates, giving also detailed insights into the ligand/proteins interaction sites.