Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is pyruvylated during 3-bromopyruvate mediated cancer cell death.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is pyruvylated during 3-bromopyruvate mediated cancer cell death.
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发表时间：
2009-12
影响因子：
2
通讯作者：
Shanmugasundaram Ganapathy-Kanniappan;J. Geschwind;Rani Kunjithapatham;Manon Buijs;J. Vossen;Irina Tchernyshyov;R. Cole;Labiq H. Syed;P. Rao;S. Ota;Mustafa Vali
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作者：
Shanmugasundaram Ganapathy-Kanniappan;J. Geschwind;Rani Kunjithapatham;Manon Buijs;J. Vossen;Irina Tchernyshyov;R. Cole;Labiq H. Syed;P. Rao;S. Ota;Mustafa Vali
背景技术丙酮酸类似物3-溴丙酮酸(3BrPA)是一种烷化剂，已知其通过阻断糖酵解来诱导癌细胞死亡。 3BrPA 的抗糖酵解作用被认为是使糖酵解酶失活。然而，缺乏关于 3BrPA 在其抗癌作用过程中与任何建议靶点直接相互作用的实验记录。方法和结果在本研究中，在多种癌细胞系中使用放射性标记的 ((14)C) 3BrPA，基于二维 (2D) 凝胶电泳放射自显影、质谱和免疫沉淀，将甘油醛-3-磷酸脱氢酶 (GAPDH) 确定为 3BrPA 的主要细胞内靶标。此外，体外酶动力学研究证实 3BrPA 对 GAPDH 具有显着的亲和力。最后，Annexin V 染色和活性 caspase-3 免疫印迹证明 3BrPA 诱导细胞凋亡。结论 3BrPA 对 GAPDH 的丙酮酰化影响其酶功能，并且是 3BrPA 介导的癌细胞死亡的主要细胞内靶标。
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.