A Cell-Potent Bisubstrate Inhibitor to Probe NatD Acetyltransferase Activity

IF 3.8 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yi-Hsun Ho, , , Emma K. Seipp, , , Thitiwat Larndate, , and , Rong Huang*, 
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Abstract

N-terminal acetyltransferase D (NatD) is a highly selective enzyme that acetylates the α-N-terminal amine of histones H4 and H2A, which share the SGRGK motif. Elevated NatD expression has been observed in lung, colorectal, breast, and bone cancer tissues, and is correlated with poor patient survival in these cancer types. In non-small cell lung cancer, NatD depletion reduces progression by repressing the epithelial-to-mesenchymal transition (EMT). Hence, NatD is a potential epigenetic target for lung cancer. To unravel the functions of NatD, a cell-potent and selective NatD inhibitor is needed to investigate the acetyltransferase activity of NatD in cancer progression. We previously reported potent and selective NatD bisubstrate inhibitors, designed by covalently linking coenzyme A to peptide substrates via an acetyl and propionyl linker. However, these inhibitors are not cell-active, limiting their application for cellular studies. Here, we designed cell-permeable bisubstrate analogs by attaching cell-penetrating peptides (CPP) to the bisubstrate inhibitor. The inhibitor displayed a Ki value of 23 nM and effectively reduced cellular Nα-acetylation on histone H4, leading to reduced migration in lung cancer cells by modulating the expression of E-cadherin, N-cadherin, and vimentin. Our findings demonstrate that the CPP-conjugated NatD inhibitor serves as a valuable chemical probe for elucidating the biological roles of NatD in lung cancer, laying the groundwork for future therapeutic strategies targeting NatD.

Abstract Image

一种检测NatD乙酰转移酶活性的细胞有效双底物抑制剂。
n -末端乙酰转移酶D (N-terminal acetyltransferase D, NatD)是一种高度选择性的酶,可使具有SGRGK基序的组蛋白H4和H2A的α- n -末端胺乙酰化。在肺癌、结直肠癌、乳腺癌和骨癌组织中观察到NatD表达升高,并与这些癌症类型中较差的患者生存率相关。在非小细胞肺癌中,NatD消耗通过抑制上皮细胞到间质转化(EMT)来减少进展。因此,NatD是肺癌的潜在表观遗传靶点。为了揭示NatD的功能,需要一种细胞有效的、选择性的NatD抑制剂来研究NatD在癌症进展中的乙酰转移酶活性。我们之前报道过有效和选择性的NatD双底物抑制剂,通过乙酰基和丙基连接剂将辅酶A共价连接到肽底物上。然而,这些抑制剂不具有细胞活性,限制了它们在细胞研究中的应用。在这里,我们通过将细胞穿透肽(CPP)连接到双底物抑制剂上,设计了具有细胞渗透性的双底物类似物。该抑制剂Ki值为23 nM,可通过调节E-cadherin、N-cadherin和vimentin的表达,有效降低组蛋白H4上n α-乙酰化,从而减少肺癌细胞的迁移。我们的研究结果表明,cpp偶联的NatD抑制剂可以作为一种有价值的化学探针来阐明NatD在肺癌中的生物学作用,为未来针对NatD的治疗策略奠定基础。
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来源期刊
ACS Chemical Biology
ACS Chemical Biology 生物-生化与分子生物学
CiteScore
7.50
自引率
5.00%
发文量
353
审稿时长
3.3 months
期刊介绍: ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology. The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies. We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
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