A Dual‐Response DNA Probe for Simultaneously Monitoring Enzymatic Activity and Environmental pH Using a Nanopore. Issue 42 (5th September 2019)

• Record Type: Journal Article
• Authors: Liu, Lei; You, Yi; Zhou, Ke; Guo, Bingyuan; Cao, Zhong; Zhao, Yuliang; Wu, Hai‐Chen
• Is Part Of: Angewandte Chemie international edition. Volume 58:Issue 42(2019)
• Page Start: 14929
• Page End: 14934
• Publication Date: 2019-09-05
• Subjects: DNA probe -- host–guest systems -- nanopore -- simultaneous quantification -- single-molecule studies; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201907816
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Both protease overexpression and local pH changes are key signatures of cancer. However, the sensitive detection of protease activities and the accurate measurement of pH in a tumor environment remain challenging. Here, we develop a dual‐response DNA probe that can simultaneously monitor protease activities and measure the local pH by translocation through α‐hemolysin (αHL). The DNA probe bears a short peptide containing phenylalanine at a pre‐designed position. Enzymatic cleavage of the peptide either exposes or removes the N‐terminal phenylalanine that can form a complex with cucurbit[7]uril. Translocation of the DNA hybrid through αHL generates current signatures that can be used to quantify protease activities. Furthermore, the current signatures possess a pH‐dependent pattern that reflects the local pH. Our results demonstrate that the versatile DNA probe may be further explored for simultaneously measuring multiple parameters of a complex system such as single cells in the future. Abstract : Two functions in one nanopore : Enzymatic cleavage of a peptide on a DNA probe either exposes or removes an N‐terminal phenylalanine that can form a complex with cucurbit[7]uril. Translocation of the DNA hybrid through the α‐hemolysin nanopore generates current signatures that can be used to quantify protease activity. The current signature also reflects the pH of the environment.