Lysine l-lactylation is the dominant lactylation isomer induced by glycolysis
Di Zhang, Jinjun Gao, Zhijun Zhu, Qian‐Ying Mao, Zhiqiang Xu, Pankaj Kumar Singh, Cornelius Rimayi, Carlos Moreno–Yruela, Shuling Xu, Gongyu Li, Yi-Cheng Sin, Yue Chen, Christian A. Olsen, Nathaniel W. Snyder, Lunzhi Dai, Lingjun Li, Yingming Zhao
Abstract
Abstract 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.