Multiplexed single-molecule enzyme activity analysis for counting disease-related proteins in biological samples
Multiplexed single-molecule enzyme activity analysis for counting disease-related proteins in biological samples
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DOI：
10.1126/sciadv.aay0888
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发表时间：
2020-03-01
期刊：
影响因子：
13.6
通讯作者：
Urano, Yasuteru
中科院分区：
文献类型：
--
作者：
Sakamoto, Shingo;Komatsu, Toru;Urano, Yasuteru
我们建立了一个超灵敏的方法，通过使用多路复用的微器件为基础的单分子酶测定来识别生物样品中的多种酶。我们使用了一个范例，其中我们“计数”的酶分子的数量，通过分析他们的单一酶活性特性对多个底物。在这项基于单酶活性的蛋白质谱（SEAP）的概念验证研究中，我们能够在单分子水平上以亚型区分方式检测血液样品中各种磷酸酯水解酶（如碱性磷酸酶、酪氨酸磷酸酶和外核苷酸焦磷酸酶）的活性，证明了其在基于酶的超灵敏检测的疾病诊断中的潜在有用性。
We established an ultrasensitive method for identifying multiple enzymes in biological samples by using a multiplexed microdevice-based single-molecule enzymatic assay. We used a paradigm in which we "count" the number of enzyme molecules by profiling their single enzyme activity characteristics toward multiple substrates. In this proof-of-concept study of the single enzyme activity-based protein profiling (SEAP), we were able to detect the activities of various phosphoric ester-hydrolyzing enzymes such as alkaline phosphatases, tyrosine phosphatases, and ectonucleotide pyrophosphatases in blood samples at the single-molecule level and in a subtype-discriminating manner, demonstrating its potential usefulness for the diagnosis of diseases based on ultrasensitive detection of enzymes.