费斯特共振能量转移
蛋白酵素
DNA
蛋白酶
肽
生物物理学
化学
生物
生物化学
酶
荧光
物理
量子力学
作者
Hieu Bui,Carl W. Brown,Susan Buckhout‐White,Sebastián A. Dı́az,Michael H. Stewart,Kimihiro Susumu,Eunkeu Oh,Mario G. Ancona,Ellen R. Goldman,Igor L. Medintz
出处
期刊:Small
[Wiley]
日期:2019-02-25
卷期号:15 (14)
被引量:17
标识
DOI:10.1002/smll.201805384
摘要
Abstract DNA can process information through sequence‐based reorganization but cannot typically receive input information from most biological processes and translate that into DNA compatible language. Coupling DNA to a substrate responsive to biological events can address this limitation. A two‐component sensor incorporating a chimeric peptide‐DNA substrate is evaluated here as a protease‐to‐DNA signal convertor which transduces protease activity through DNA gates that discriminate between different input proteases. Acceptor dye‐labeled peptide‐DNAs are assembled onto semiconductor quantum dot (QD) donors as the input gate. Addition of trypsin or chymotrypsin cleaves their cognate peptide sequence altering the efficiency of Förster resonance energy transfer (FRET) with the QD and frees a DNA output which interacts with a tetrahedral output gate. Downstream output gate rearrangement results in FRET sensitization of a new acceptor dye. Following characterization of component assembly and optimization of individual steps, sensor ability to discriminate between the two proteases is confirmed along with effects from joint interactions where potential for cross‐talk is highest. Processing multiple bits of information for a sensing outcome provides more confidence than relying on a single change especially for the discrimination between different targets. Coupling other substrates to DNA that respond similarly could help target other types of enzymes.
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