Department of Medicinal Chemistry and Molecular Pharmacology, Purdue Center for Cancer Research, Purdue University, 575 Stadium Mall Drive, West Lafayette, IN, 47905, USA.
Angew Chem Int Ed Engl. 2016 Aug 8;55(33):9562-6. doi: 10.1002/anie.201603387. Epub 2016 Jun 29.
A sensing approach is applied to encode quantitative enzymatic activity information into DNA sequence populations. The method utilizes DNA-linked peptide substrates as activity probes. Signal detection involves chemical manipulation of a probe population downstream of sample exposure and application of purifying, selective pressure for enzyme products. Selection-induced changes in DNA abundance indicate sample activity. The detection of protein kinase, protease, and farnesyltransferase activities is demonstrated. The assays were employed to measure enzyme inhibition by small molecules and activity in cell lysates using parallel DNA sequencing or quantitative PCR. This strategy will allow the extensive infrastructure for genetic analysis to be applied to proteomic assays, which has a number of advantages in throughput, sensitivity, and sample multiplexing.
一种传感方法被应用于将定量酶活性信息编码到 DNA 序列群体中。该方法利用 DNA 连接的肽底物作为活性探针。信号检测涉及在样本暴露后对探针群体进行化学处理，并对酶产物进行纯化、选择性压力。DNA 丰度的选择诱导变化表明了样品的活性。本文展示了蛋白激酶、蛋白酶和法呢基转移酶活性的检测。这些测定方法被用于使用平行 DNA 测序或定量 PCR 来测量小分子对酶的抑制作用和细胞裂解物中的活性。该策略将使广泛的遗传分析基础设施应用于蛋白质组学分析，这在通量、灵敏度和样品多路复用方面具有许多优势。
