核糖核苷酸,广泛存在于所有生物体中,对生理和病理过程都至关重要,可以自然出现为核糖核苷单,di-,和三磷酸盐。天然核糖核苷酸也可以在不同的磷酸化形式之间动态切换,对感知构成了巨大挑战。专门设计的纳米孔传感器有望完全区分所有规范的核糖核苷单,di-,和三磷酸盐。然而,这种示威从未被报道过,由于缺乏具有足够分辨率的合适的纳米孔传感器。在这项工作中,我们利用苯基硼酸(PBA)修饰的耻垢分枝杆菌孔蛋白A(MspA)异八聚体进行核糖核苷酸传感。十二种核糖核苷酸,包括mono-,di-,和胞苷三磷酸盐(CMP,CDP,CTP),尿苷(UMP,UDP,UTP),腺苷(AMP,ADP,ATP),和鸟苷(GMP,GDP,GTP)同时进行鉴别。还开发了一种机器学习算法,其对于核糖核苷酸感测实现了99.9%的一般准确度。该策略还进一步应用于鉴定ATP片剂和注射剂中的核糖核苷酸组分。这种感知策略提供了一种直接的,准确,easy,和快速解决方案,以表征不同磷酸化形式的核糖核苷酸成分。
Ribonucleotides, which widely exist in all living organisms and are essential to both physiological and pathological processes, can naturally appear as ribonucleoside mono-, di-, and triphosphates. Natural
ribonucleotides can also dynamically switch between different phosphorylated forms, posing a great challenge for sensing. A specially engineered nanopore sensor is promising for full discrimination of all canonical ribonucleoside mono-, di-, and triphosphates. However, such a demonstration has never been reported, due to the lack of a suitable nanopore sensor that has a sufficient resolution. In this work, we utilized a phenylboronic acid (PBA) modified Mycobacterium smegmatis porin A (MspA) hetero-octamer for ribonucleotide sensing. Twelve types of
ribonucleotides, including mono-, di-, and triphosphates of cytidine (CMP, CDP, CTP), uridine (UMP, UDP, UTP), adenosine (AMP, ADP, ATP), and guanosine (GMP, GDP, GTP) were simultaneously discriminated. A machine-learning algorithm was also developed, which achieved a general accuracy of 99.9% for ribonucleotide sensing. This strategy was also further applied to identify ribonucleotide components in ATP tablets and injections. This sensing strategy provides a direct, accurate, easy, and rapid solution to characterize ribonucleotide components in different phosphorylated forms.