Lysine l-lactylation (K_l-la) is a novel protein posttranslational modification (PTM) driven by l-lactate. This PTM has three isomers: K_l-la, N-ε-(carboxyethyl)-lysine (K_ce) and d-lactyl-lysine (K_d-la), which are often confused in the context of the Warburg effect and nuclear presence. Here we introduce two methods to differentiate these isomers: a chemical derivatization and high-performance liquid chromatography analysis for efficient separation, and isomer-specific antibodies for high-selectivity identification. We demonstrated that K_l-la is the primary lactylation isomer on histones and dynamically regulated by glycolysis, not K_d-la or K_ce, which are observed when the glyoxalase system was incomplete. The study also reveals that lactyl-coenzyme A, a precursor in l-lactylation, correlates positively with K_l-la levels. This work not only provides a methodology for distinguishing other PTM isomers, but also highlights K_l-la as the primary responder to glycolysis and the Warburg effect.

赖氨酸L-乳酰化(Kl _{- la} )是一种由L-乳酸驱动的新型蛋白质翻译后修饰(PTM)。该 PTM 具有三种异构体：K _{l -la} 、 N -ε-(羧乙基)-赖氨酸 (K _ce ) 和d -乳酰-赖氨酸 (K _{d -la} )，它们在瓦尔堡效应和核反应中经常被混淆。在场。在这里，我们介绍两种区分这些异构体的方法：化学衍生化和高效液相色谱分析以进行有效分离，以及异构体特异性抗体以进行高选择性鉴定。我们证明，K _{l -la}是组蛋白上的主要乳酰化异构体，并受糖酵解动态调节，而不是当乙二醛酶系统不完整时观察到的 K _{d -la}或 K _ce 。该研究还表明， L-乳酰化的前体乳酰辅酶 A 与K _-la水平呈正相关。这项工作不仅提供了区分其他 PTM 异构体的方法，而且还强调了 K _{l -la}作为糖酵解和 Warburg 效应的主要反应物。