3-Bromopyruvate (3BP) is a potential anticancer agent viewed as a glycolytic inhibitor that preferentially kills cancer cells through inhibition of glyceraldehyde 3-phosphate dehydrogenase (GAPDH), resulting in severe energy depletion. We previously identified four cysteine residues in GAPDH that are alkylated by 3BP, resulting in its inactivation. However, we also showed that addition of excess pyruvate, the final product of glycolysis, was unable to rescue cells from 3BP treatment. This result indicates that GAPDH may not be the only relevant target and is consistent with the chemical reactivity of 3BP that should result in the modification of cysteine residues in many different proteins. To directly test this hypothesis, we first synthesized a probe of 3BP activity bearing an alkyne functionality, termed AO3BP, and then demonstrated that this probe could modify a variety of proteins in living cells. Subsequent competition of AO3BP labeling with pretreatment by 3BP identified 62 statistically significant proteins of various functions as targets of 3BP, confirming that 3BP labeling is indeed widespread. We conclude that 3BP’s cytotoxic impact on cancer cells does not only result from selective inhibition of glycolysis but rather from a more widespread effect on cellular proteins that could be driven by the pharmacokinetics of the 3BP. These pleiotropic consequences should be considered when thinking about the potential toxicity of this highly reactive compound.

中文翻译：

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癌细胞蛋白质组的生物正交分析确定了糖酵解以外的大量3-Bromopyruvate目标

3-溴丙酮酸酯（3BP）是一种潜在的抗癌剂，被认为是一种糖酵解抑制剂，可通过抑制3-磷酸甘油醛脱氢酶（GAPDH）优先杀死癌细胞，从而导致严重的能量消耗。我们之前在GAPDH中发现了4个被3BP烷基化的半胱氨酸残基，从而使其失活。但是，我们还表明，添加过量的丙酮酸（糖酵解的最终产物）无法从3BP处理中拯救细胞。该结果表明，GAPDH可能不是唯一的相关靶标，并且与3BP的化学反应性一致，后者应导致许多不同蛋白质中半胱氨酸残基的修饰。为了直接验证这一假设，我们首先合成了带有炔烃功能的3BP活性探针，称为AO3BP，然后证明该探针可以修饰活细胞中的多种蛋白质。AO3BP标记与3BP预处理的竞争随后鉴定出62种具有各种功能的统计学上显着的蛋白质作为3BP的靶标，从而证实3BP标记的确广泛存在。我们得出的结论是3BP对癌细胞的细胞毒性影响不仅是由于糖酵解的选择性抑制所致，而且还归因于对3BP药代动力学可能驱动的细胞蛋白质的更广泛影响。考虑这种高反应性化合物的潜在毒性时，应考虑这些多效性后果。AO3BP标记与3BP预处理的竞争随后鉴定出62种具有各种功能的统计学上显着的蛋白质作为3BP的靶标，从而证实3BP标记的确是广泛存在的。我们得出的结论是3BP对癌细胞的细胞毒性影响不仅是由于糖酵解的选择性抑制所致，而且还归因于对3BP药代动力学可能驱动的细胞蛋白质的更广泛影响。考虑这种高反应性化合物的潜在毒性时，应考虑这些多效性后果。AO3BP标记与3BP预处理的竞争随后鉴定出62种具有各种功能的统计学上显着的蛋白质作为3BP的靶标，从而证实3BP标记的确是广泛存在的。我们得出的结论是3BP对癌细胞的细胞毒性影响不仅是由于糖酵解的选择性抑制所致，而且还归因于对3BP药代动力学可能驱动的细胞蛋白质的更广泛影响。考虑这种高反应性化合物的潜在毒性时，应考虑这些多效性后果。我们得出的结论是3BP对癌细胞的细胞毒性影响不仅是由于糖酵解的选择性抑制所致，而且还归因于对3BP药代动力学可能驱动的细胞蛋白质的更广泛影响。考虑这种高反应性化合物的潜在毒性时，应考虑这些多效性后果。我们得出的结论是3BP对癌细胞的细胞毒性影响不仅是由于糖酵解的选择性抑制所致，而且还归因于对3BP药代动力学可能驱动的细胞蛋白质的更广泛影响。考虑这种高反应性化合物的潜在毒性时，应考虑这些多效性后果。