%0 Journal Article %A SHANMUGASUNDARAM GANAPATHY-KANNIAPPAN %A JEAN-FRANCOIS H. GESCHWIND %A RANI KUNJITHAPATHAM %A MANON BUIJS %A JOSEPHINA A. VOSSEN %A IRINA TCHERNYSHYOV %A ROBERT N. COLE %A LABIQ H. SYED %A PRAMOD P. RAO %A SHINICHI OTA %A MUSTAFA VALI %T Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Is Pyruvylated during 3-Bromopyruvate Mediated Cancer Cell Death %D 2009 %J Anticancer Research %P 4909-4918 %V 29 %N 12 %X Background: The pyruvic acid analog 3-bromopyruvate (3BrPA) is an alkylating agent known to induce cancer cell death by blocking glycolysis. The anti-glycolytic effect of 3BrPA is considered to be the inactivation of glycolytic enzymes. Yet, there is a lack of experimental documentation on the direct interaction of 3BrPA with any of the suggested targets during its anticancer effect. Methods and Results: In the current study, using radiolabeled (14C) 3BrPA in multiple cancer cell lines, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as the primary intracellular target of 3BrPA, based on two-dimensional (2D) gel electrophoretic autoradiography, mass spectrometry and immunoprecipitation. Furthermore, in vitro enzyme kinetic studies established that 3BrPA has marked affinity to GAPDH. Finally, Annexin V staining and active caspase-3 immunoblotting demonstrated that apoptosis was induced by 3BrPA. Conclusion: GAPDH pyruvylation by 3BrPA affects its enzymatic function and is the primary intracellular target in 3BrPA mediated cancer cell death. %U https://ar.iiarjournals.org/content/anticanres/29/12/4909.full.pdf