As enzymes that mediate the attachment of long-chain fatty acids to cysteine residues, ZDHHC proteins have been reported to be promising therapeutic targets for treating cancer and autoimmune diseases. Yet, due to the lack of potent selective inhibitors, scrutiny of the biological functions of ZDHHCs has been limited. The main hindrance for developing ZDHHC inhibitors is the lack of a facile high-throughput assay. Here, we developed a ZDHHC3/7/20 high-throughput assay based on the acylation-coupled lipophilic induction of polarization (Acyl-cLIP) method and screened several potential ZDHHC inhibitors. Furthermore, we demonstrated that in vitro results from the Acyl-cLIP assay are supported by the results from cell-based assays. We envision that this new ZDHHC3/7/20 Acyl-cLIP assay will accelerate the high-throughput screening of large compound libraries for improved ZDHHC inhibitors and provide therapeutic benefits for cancer and autoimmune diseases.

中文翻译：

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用于筛选 ZDHHC3/7/20 酰基转移酶抑制剂的高通量酶测定

作为介导长链脂肪酸与半胱氨酸残基结合的酶，ZDHHC 蛋白据报道是治疗癌症和自身免疫性疾病的有希望的治疗靶点。然而，由于缺乏有效的选择性抑制剂，对 ZDHHC 的生物学功能的审查受到了限制。开发 ZDHHC 抑制剂的主要障碍是缺乏简便的高通量检测方法。在这里，我们开发了一种基于酰化偶联亲脂极化诱导 (Acyl-cLIP) 方法的 ZDHHC3/7/20 高通量检测方法，并筛选了几种潜在的 ZDHHC 抑制剂。此外，我们证明了来自 Acyl-cLIP 测定的体外结果得到基于细胞的测定结果的支持。