Gonzatti Michelangelo B, Hintzen Jordi C J, Sharma Isha, Najar Mohd Altaf, Tsusaka Takeshi, Marcinkiewicz Mariola M, Da Silva Crispim Claudia Veronica, Snyder Nathaniel W, Burslem George M, Goldberg Emily L
Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
J Biol Chem. 2025 Aug 18:110602. doi: 10.1016/j.jbc.2025.110602.
Metabolism and post-translational modifications (PTMs) are intrinsically linked and the number of identified metabolites that can covalently modify proteins continues to increase. This metabolism/PTM crosstalk is especially true for lactate, the product of anaerobic metabolism following glycolysis. Lactate forms an amide bond with the ε-amino group of lysine, a modification known as lysine lactylation, or Kla. Multiple independent mechanisms have been proposed in the formation of Kla, including p300/CBP-dependent transfer from lactyl-CoA, a reactive intermediate lactoylglutathione species that non-enzymatically lactylates proteins, and several enzymes are reported to have lactyl transferase capability. We recently discovered that class I histone deacetylases (HDACs) 1, 2, and 3 can all reverse their canonical chemical reaction to catalyze lysine β-hydroxybutyrylation. Here we tested the hypothesis that HDACs can also catalyze Kla formation. Using biochemical, pharmacological, and genetic approaches, we found that HDACs are sufficient to catalyze Kla formation and that HDACs are a major driver of lysine lactylation. Dialysis experiments confirm this is a reversible reaction that depends on lactate concentration. We also directly quantified intracellular lactyl-CoA and found that Kla abundance can be uncoupled from lactyl-CoA levels. Therefore, we propose a model in which the majority of Kla is formed through enzymatic addition of lactate by HDACs 1, 2, and 3.
新陈代谢与翻译后修饰（PTMs）有着内在联系，能够共价修饰蛋白质的已鉴定代谢物数量持续增加。这种新陈代谢/PTM串扰在乳酸（糖酵解后无氧代谢的产物）方面尤为明显。乳酸与赖氨酸的ε-氨基形成酰胺键，这种修饰被称为赖氨酸乳酰化，即Kla。关于Kla的形成已提出多种独立机制，包括依赖p300/CBP从乳酰辅酶A进行转移、一种能非酶促地使蛋白质乳酰化的活性中间产物乳酰谷胱甘肽物种，并且有报道称几种酶具有乳酰转移酶能力。我们最近发现I类组蛋白去乙酰化酶（HDACs）1、2和3都能逆转其经典化学反应以催化赖氨酸β-羟基丁酰化。在此，我们测试了HDACs也能催化Kla形成这一假说。通过生化、药理学和遗传学方法，我们发现HDACs足以催化Kla形成，并且HDACs是赖氨酸乳酰化的主要驱动因素。透析实验证实这是一个依赖乳酸浓度的可逆反应。我们还直接定量了细胞内的乳酰辅酶A，发现Kla丰度可与乳酰辅酶A水平解偶联。因此，我们提出了一个模型，其中大部分Kla是由HDACs 1、2和3通过酶促添加乳酸形成的。
