A palmitoyl transferase chemical-genetic system to map ZDHHC-specific S-acylation
A palmitoyl transferase chemical-genetic system to map ZDHHC-specific S-acylation
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DOI：
10.1038/s41587-023-02030-0
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发表时间：
2024-01-08
影响因子：
46.9
通讯作者：
Tate，Edward W.
中科院分区：
文献类型：
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作者：
Ocasio，Cory A.;Baggelaar，Marc P.;Tate，Edward W.
23种含Asp-His-His-Cys基序的人锌指（ZDHHC）S-酰基转移酶催化半胱氨酸残基处的长链S-酰基化，所述长链S-酰基化跨越对正常生理学重要或在疾病中失调的数百种蛋白质的广泛网络。在这里，我们提出了一种技术，直接映射的蛋白质底物的特定ZDHHC在整个蛋白质组水平，在完整的细胞。成对的ZDHHC“空穴”突变体和“凸起的”化学标记的脂肪酸探针的结构引导工程使探针能够转移到特异性蛋白质底物，具有优于野生型ZDHHC的选择性。化学遗传学系统以5种人ZDHHC（3、7、11、15和20）为例，并应用于产生从头ZDHHC底物谱，在多个细胞系中鉴定出>300种底物和新的功能多样的蛋白质的S-酰化位点。我们期望这个平台将阐明广泛的模型和生物体的S-酰化生物学。
The 23 human zinc finger Asp-His-His-Cys motif-containing (ZDHHC)S-acyltransferases catalyze long-chainS-acylation at cysteine residues across an extensive network of hundreds of proteins important for normal physiology or dysregulated in disease. Here we present a technology to directly map the protein substrates of a specific ZDHHC at the whole-proteome level, in intact cells. Structure-guided engineering of paired ZDHHC ‘hole’ mutants and ‘bumped’ chemically tagged fatty acid probes enabled probe transfer to specific protein substrates with excellent selectivity over wild-type ZDHHCs. Chemical–genetic systems were exemplified for five human ZDHHCs (3, 7, 11, 15 and 20) and applied to generate de novo ZDHHC substrate profiles, identifying >300 substrates andS-acylation sites for new functionally diverse proteins across multiple cell lines. We expect that this platform will elucidateS-acylation biology for a wide range of models and organisms.