Harnessing Ionic Selectivity in Acetyltransferase Chemoproteomic Probes,ACS Chemical Biology

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Harnessing Ionic Selectivity in Acetyltransferase Chemoproteomic Probes
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2020-12-29 , DOI: 10.1021/acschembio.0c00766
Yihang Jing ₁ , Jose L Montano ₁ , Michaella Levy ₂ , Jeffrey E Lopez ₁ , Pei-Pei Kung ₃ , Paul Richardson ₃ , Krzysztof Krajewski ₄ , Laurence Florens ₂ , Michael P Washburn _{2,

5} , Jordan L Meier ₁

Affiliation

Chemical proteomics provides a powerful strategy for the high-throughput assignment of enzyme function or inhibitor selectivity. However, identifying optimized probes for an enzyme family member of interest and differentiating signal from the background remain persistent challenges in the field. To address this obstacle, here we report a physiochemical discernment strategy for optimizing chemical proteomics based on the coenzyme A (CoA) cofactor. First, we synthesize a pair of CoA-based sepharose pulldown resins differentiated by a single negatively charged residue and find this change alters their capture properties in gel-based profiling experiments. Next, we integrate these probes with quantitative proteomics and benchmark analysis of “probe selectivity” versus traditional “competitive chemical proteomics.” This reveals that the former is well-suited for the identification of optimized pulldown probes for specific enzyme family members, while the latter may have advantages in discovery applications. Finally, we apply our anionic CoA pulldown probe to evaluate the selectivity of a recently reported small molecule N-terminal acetyltransferase inhibitor. These studies further validate the use of physical discriminant strategies in chemoproteomic hit identification and demonstrate how CoA-based chemoproteomic probes can be used to evaluate the selectivity of small molecule protein acetyltransferase inhibitors, an emerging class of preclinical therapeutic agents.

中文翻译：

利用乙酰转移酶化学蛋白质组探针的离子选择性

化学蛋白质组学为酶功能或抑制剂选择性的高通量分配提供了强大的策略。然而，识别感兴趣的酶家族成员的优化探针以及从背景中区分信号仍然是该领域持续存在的挑战。为了解决这个障碍，我们在这里报告了一种基于辅酶 A (CoA) 辅因子来优化化学蛋白质组学的理化识别策略。首先，我们合成了一对由单个带负电荷残基区分的基于 CoA 的琼脂糖下拉树脂，并发现这种变化改变了它们在基于凝胶的分析实验中的捕获特性。接下来，我们将这些探针与定量蛋白质组学以及“探针选择性”与传统“竞争性化学蛋白质组学”的基准分析相结合。这表明前者非常适合识别特定酶家族成员的优化下拉探针，而后者在发现应用中可能具有优势。最后，我们应用阴离子 CoA 下拉探针来评估最近报道的小分子 N 末端乙酰转移酶抑制剂的选择性。这些研究进一步验证了物理判别策略在化学蛋白质组命中鉴定中的应用，并证明了如何使用基于 CoA 的化学蛋白质组探针来评估小分子蛋白质乙酰转移酶抑制剂（一类新兴的临床前治疗药物）的选择性。

更新日期：2021-01-15

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