Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is pyruvylated during 3-bromopyruvate mediated cancer cell death

Shanmugasundaram Ganapathy-Kanniappan, Jean Francois H. Geschwind, Rani Kunjithapatham, Manon Buijs, Josephina A. Vossen, Irina Tchernyshyov, Robert N. Cole, Labiq H. Syed, Pramod P. Rao, Shinichi Ota, Mustafa Vali

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)4909-4918
Number of pages10
JournalAnticancer research
Volume29
Issue number12
StatePublished - Dec 2009

Keywords

  • 3-Bromopyruvate
  • Cancer cell death
  • GAPDH
  • Glycolysis
  • Hep3B
  • HepG2
  • SK-Hepl

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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