Lysine lipoylation plays vital roles in cell metabolism and redox processes. For example, removal of lipoylation will decrease pyruvate dehydrogenase activity and affect the citric acid cycle. Despite the important functions of lysine lipoylation, the mechanisms for the addition and removal of this modification remain largely unexplored. Very few useful chemical tools are available to study the interactions of lysine lipoylation with its regulatory delipoylation proteins. For example, immunoaffinity purification-mass spectrometry is one of such tools, which highly relies on antibody efficiency and purification techniques. Single-step activity based fluorogenic probes developed by our groups and others is also an efficient method to study the deacylation activity. Affinity-based labeling probe using photo-cross-linker is a powerful platform to study the transient and dynamic interactions of peptide ligands with the interacting proteins. Herein, we have designed and synthesized a dual-function probe KTLlip for studying enzymatic delipoylation (eraser) activity and interaction of lysine lipoylation with the eraser at the same time. We show that KTLlip can be used as a useful tool to detect delipoylation as demonstrated by its ability to fluorescently label the eraser activity of recombinant Sirt2. We envision that the probe will help delineate the roles of delipoylation enzyme in biology.

赖氨酸脂酰化在细胞代谢和氧化还原过程中起着至关重要的作用。例如，去除脂酰化将降低丙酮酸脱氢酶活性并影响柠檬酸循环。尽管赖氨酸脂酰化具有重要功能，但添加和去除这种修饰的机理仍未得到充分探索。很少有有用的化学工具可用来研究赖氨酸脂酰化及其调节性脱脂蛋白的相互作用。例如，免疫亲和纯化-质谱法是这样的工具之一，其高度依赖于抗体效率和纯化技术。我们小组和其他人开发的基于单步活性的荧光探针也是研究脱酰活性的有效方法。使用光交联剂的基于亲和力的标记探针是研究肽配体与相互作用蛋白的瞬时和动态相互作用的强大平台。本文中，我们设计并合成了一种双功能探针KTLlip，用于同时研究酶促脱脂作用（橡皮擦）和赖氨酸脂化作用与橡皮擦的相互作用。我们显示KTLlip可以用作检测脱脂作用的有用工具，其荧光标记重组Sirt2的橡皮擦活性的能力证明了这一点。我们设想该探针将有助于描述脱脂酶在生物学中的作用。我们设计并合成了一种双功能探针KTLlip，用于同时研究酶促脱脂作用（橡皮擦）和赖氨酸脂化作用与橡皮擦的相互作用。我们显示KTLlip可以用作检测脱脂作用的有用工具，其荧光标记重组Sirt2的橡皮擦活性的能力证明了这一点。我们设想该探针将有助于描述脱脂酶在生物学中的作用。我们设计并合成了一种双功能探针KTLlip，用于同时研究酶促脱脂作用（橡皮擦）和赖氨酸脂化作用与橡皮擦的相互作用。我们显示KTLlip可以用作检测脱脂作用的有用工具，其荧光标记重组Sirt2的橡皮擦活性的能力证明了这一点。我们设想该探针将有助于描述脱脂酶在生物学中的作用。