Russell H. Morgan Department of Radiology and Radiological Sciences, Baltimore, MD 21287, USA.
Anticancer Res. 2009 Dec;29(12):4909-18.
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.
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.
GAPDH pyruvylation by 3BrPA affects its enzymatic function and is the primary intracellular target in 3BrPA mediated cancer cell death.
丙酮酸类似物 3-溴丙酮酸（3BrPA）是一种烷化剂，通过阻断糖酵解诱导癌细胞死亡。3BrPA 的抗糖酵解作用被认为是使糖酵解酶失活。然而，在其抗癌作用过程中，缺乏 3BrPA 与任何建议靶点直接相互作用的实验文献。
在本研究中，使用放射性标记的（14C）3BrPA 在多种癌细胞系中，根据二维（2D）凝胶电泳放射自显影、质谱和免疫沉淀，甘油醛-3-磷酸脱氢酶（GAPDH）被鉴定为 3BrPA 的主要细胞内靶标。此外，体外酶动力学研究表明，3BrPA 对 GAPDH 具有显著的亲和力。最后，Annexin V 染色和活性 caspase-3 免疫印迹表明，凋亡是由 3BrPA 诱导的。
3BrPA 对 GAPDH 的丙酮酸化作用影响其酶功能，是 3BrPA 介导的癌细胞死亡的主要细胞内靶标。
