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

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 ((14)C) 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.

• Authors: 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
• Journal: Anticancer research (Anticancer Res) Vol. 29 Issue 12 Pg. 4909-18 (Dec 2009) ISSN: 1791-7530 [Electronic] Greece
• PMID: 20044597 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Pyruvates
• bromopyruvate
• Glyceraldehyde-3-Phosphate Dehydrogenases

Topics

• Animals
• Apoptosis (drug effects, physiology)
• Carcinoma, Hepatocellular (drug therapy, enzymology, pathology)
• Electrophoresis, Gel, Two-Dimensional
• Glyceraldehyde-3-Phosphate Dehydrogenases (metabolism)
• Humans
• Immunoblotting
• Immunoprecipitation
• Liver Neoplasms (drug therapy, enzymology, pathology)
• Pyruvates (pharmacology)
• Rabbits
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Tumor Cells, Cultured