Unique ssDNA-Induced Fluorescence Enhancement of a Conjugated Polymer Brush for Label-Free Sensing of S1 Nuclease and ATP

Nuclease plays a vital role in DNA replication, repair, and recombination. Taking advantage of the unique ssDNA-induced fluorescence enhancement of a cationic conjugated polymer brush (PPE-PLL), we constructed a new simple strategy for the detection of S1 nuclease and ATP simultaneously. It was found that the fluorescence of PPE-PLL can be enhanced by ssDNA significantly, which is quite different from traditional conjugated polymers. However, in the presence of S1 nuclease, the ssDNA was cleaved into small oligonucleotide and mononucleotide fragments. The fragments showed much weaker electrostatic interactions with PPE-PLL, resulting in a decreased fluorescence. Furthermore, when ATP was introduced as an inhibitor of S1 nuclease, a relatively high fluorescence signal was observed. Our strategy has high specificity and selectivity, and it works well in complicated system. The limit of detection for S1 nuclease and ATP can be estimated to be 0.15 U/mL and 0.068 mM, respectively. Additionally, the mechanism of the unusual ssDNA-induced enhancement of PPE-PLL was discussed by dynamic light scattering, fluorescence, and zeta potential measurements. Therefore, this simple, sensitive, label-free, and low-cost method may be extended to the sensing of other DNA-related biomolecules.

[1]  Shu Wang,et al.  Water-soluble fluorescent conjugated polymers and their interactions with biomacromolecules for sensitive biosensors. , 2010, Chemical Society reviews.

[2]  Lijuan Ou,et al.  Hairpin loop-enhanced fluorescent copper nanoclusters and application in S1 nuclease detection. , 2018, The Analyst.

[3]  H. Park,et al.  Fluorescent S1 nuclease assay utilizing exponential strand displacement amplification. , 2019, The Analyst.

[4]  K. Morikawa,et al.  Structure and function of nucleases in DNA repair: shape, grip and blade of the DNA scissors , 2002, Oncogene.

[5]  Baoxin Li,et al.  Naked-eye sensitive detection of nuclease activity using positively-charged gold nanoparticles as colorimetric probes. , 2011, Chemical communications.

[6]  B. Liu,et al.  Water-soluble conjugated polymers as the platform for protein sensors , 2010 .

[7]  G. Bazan,et al.  Fluorescein provides a resonance gate for FRET from conjugated polymers to DNA intercalated dyes. , 2004, Journal of the American Chemical Society.

[8]  R. Yu,et al.  MnO2 Nanosheet-based Fluorescence Sensing Platform for Sensitive Detection of Endonuclease. , 2017, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[9]  K. Schanze,et al.  Solvent‐Induced Self‐Assembly of a Meta‐Linked Conjugated Polyelectrolyte. Helix Formation, Guest Intercalation, and Amplified Quenching , 2004 .

[10]  W. Qin,et al.  Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode. , 2013, Biosensors & bioelectronics.

[11]  G. Bazan,et al.  Conjugated-Polymer-Amplified Sensing, Imaging, and Therapy , 2017 .

[12]  B. Liu,et al.  Fluorescence resonance energy transfer between an anionic conjugated polymer and a dye-labeled lysozyme aptamer for specific lysozyme detection. , 2009, Chemical communications.

[13]  Shulin Zhao,et al.  A G-quadruplex-based colorimetric assay of S1 nuclease activity and inhibition , 2015 .

[14]  J. Solomon,et al.  Quantitative high-performance liquid chromatography analysis of DNA oxidized in vitro and in vivo. , 1991, Analytical biochemistry.

[15]  A. Terzic,et al.  ATP-sensitive potassium channels: metabolic sensing and cardioprotection. , 2007, Journal of applied physiology.

[16]  Xiaomiao Feng,et al.  An ultrasensitive label-free biosensor for assaying of sequence-specific DNA-binding protein based on amplifying fluorescent conjugated polymer. , 2013, Biosensors & bioelectronics.

[17]  Alexander V. Gourine,et al.  ATP is a mediator of chemosensory transduction in the central nervous system , 2005, Nature.

[18]  J. Chao,et al.  Thioflavin T as an Efficient G-Quadruplex Inducer for the Highly Sensitive Detection of Thrombin Using a New Föster Resonance Energy Transfer System. , 2015, ACS applied materials & interfaces.

[19]  Kemin Wang,et al.  Poly(thymine)-templated fluorescent copper nanoparticles for ultrasensitive label-free nuclease assay and its inhibitors screening. , 2013, Analytical chemistry.

[20]  Wei Huang,et al.  Conjugated Polymer Brush Based on Poly(l-lysine) with Efficient Ovalbumin Delivery for Dendritic Cell Vaccine. , 2018, ACS applied bio materials.

[21]  Wei Huang,et al.  An optical-logic system based on cationic conjugated polymer/DNA/intercalating dyes assembly for label-free detection of conformational conversion of DNA i-motif structure , 2011 .

[22]  G. Shen,et al.  Double-strand DNA-templated formation of copper nanoparticles as fluorescent probe for label free nuclease enzyme detection. , 2013, Biosensors & bioelectronics.

[23]  Jiaona Xu,et al.  The aptamer DNA-templated fluorescence silver nanoclusters: ATP detection and preliminary mechanism investigation. , 2017, Biosensors & bioelectronics.

[24]  Yaodong Zhang,et al.  Label-free fluorometric detection of S1 nuclease activity by using polycytosine oligonucleotide-templated silver nanoclusters. , 2015, Analytical biochemistry.

[25]  S. Gite,et al.  Single-strand-specific nucleases. , 1995, Critical reviews in microbiology.

[26]  Shulin Zhao,et al.  A sensitive fluorescence turn-on assay of bleomycin and nuclease using WS2 nanosheet as an effective sensing platform. , 2015, Analytica chimica acta.

[27]  A. Jeltsch,et al.  A fast and accurate enzyme-linked immunosorbent assay for the determination of the DNA cleavage activity of restriction endonucleases. , 1993, Analytical biochemistry.

[28]  Xinrui Duan,et al.  Cationic conjugated polymers for optical detection of DNA methylation, lesions, and single nucleotide polymorphisms. , 2010, Accounts of chemical research.

[29]  Qi Zhao,et al.  Label-Free Fluorescence Assay of S1 Nuclease and Hydroxyl Radicals Based on Water-Soluble Conjugated Polymers and WS2 Nanosheets , 2016, Sensors.

[30]  Wei Huang,et al.  DNA biosensors based on water-soluble conjugated polymers. , 2011, Biosensors & bioelectronics.

[31]  Wenli Song,et al.  Water-Soluble Iridium(III)-Containing Conjugated Polyelectrolytes with Weakened Energy Transfer Properties for Multicolor Protein Sensing Applications , 2011 .

[32]  R. Rissman,et al.  Characterization of ATP alternations in an Alzheimer's disease transgenic mouse model. , 2015, Journal of Alzheimer's disease : JAD.

[33]  Sachiko Chikahisa,et al.  The Role of ATP in Sleep Regulation , 2011, Front. Neur..

[34]  Hongtao Yu,et al.  Interface engineering catalytic graphene for smart colorimetric biosensing. , 2012, ACS nano.

[35]  E. Rangarajan,et al.  Sugar non-specific endonucleases. , 2001, FEMS microbiology reviews.

[36]  Ke Ma,et al.  A label-free aptasensor for turn-on fluorescent detection of ATP based on AIE-active probe and water-soluble carbon nanotubes , 2016 .

[37]  L. Owen,et al.  Metabolic Agents that Enhance ATP can Improve Cognitive Functioning: A Review of the Evidence for Glucose, Oxygen, Pyruvate, Creatine, and L-Carnitine , 2011, Nutrients.

[38]  B Sugden,et al.  Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. , 1973, Biochemistry.