N-Acetyl modification, a chemical modification commonly found on biomacromolecules, plays a crucial role in the regulation of cell activities and is related to a variety of diseases. However, due to the instability of N-acetyl modification, accurate and rapid identification of N-acetyl modification with a low measurement cost is still technically challenging. Here, based on hydroxylamine deacetylation and nanopore single molecule chemistry, a universal sensing strategy for N-acetyl modification has been developed. Acetohydroxamic acid (AHA), which is produced by the hydroxylamine deacetylation reaction and serves as a reporter for N-acetylation identification, is specifically sensed by a phenylboronic acid (PBA)-modified Mycobacterium smegmatis porin A (MspA). With this strategy, N-acetyl modifications on RNA, DNA, proteins, and glycans were identified, demonstrating its generality. Specifically, histones can be treated with hydroxylamine deacetylation, from which the generated AHA can represent the amount of N-acetyl modification detected by a nanopore sensor. The unique event features of AHA also demonstrate the robustness of sensing against other interfering analytes in the environment.

中文翻译：

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通过羟胺脱乙酰化进行 N-乙酰化的纳米孔鉴定 (NINAHD)

N-乙酰基修饰是生物大分子上常见的化学修饰，在细胞活动的调节中发挥着至关重要的作用，与多种疾病相关。然而，由于N-乙酰基修饰的不稳定性，以较低的测量成本准确、快速地鉴定N-乙酰基修饰仍然具有技术挑战性。在此，基于羟胺脱乙酰化和纳米孔单分子化学，开发了一种用于N-乙酰基修饰的通用传感策略。乙酰羟肟酸 (AHA) 是由羟胺脱乙酰化反应产生的，可作为 N-乙酰化鉴定的报告分子，可被苯硼酸 (PBA) 修饰的耻垢分枝杆菌孔蛋白 A (MspA) 特异性感知。通过该策略，鉴定了 RNA、DNA、蛋白质和聚糖上的N-乙酰基修饰，证明了其普遍性。具体来说，可以对组蛋白进行羟胺脱乙酰化处理，生成的AHA可以代表纳米孔传感器检测到的N-乙酰基修饰量。 AHA 独特的事件特征还证明了传感对环境中其他干扰分析物的鲁棒性。