Colorimetric and electrochemical determination of the activity of protein kinase based on retarded particle growth due to binding of phosphorylated peptides to DNA – capped silver nanoclusters

Shen, Congcong; Zhang, Kaina; Gao, Nanxing; Wei, Shijiong; Liu, Guimei; Chai, Yuanlin; Yang, Minghui

doi:10.1007/s00604-016-1944-y

Cited by 20 publications

(3 citation statements)

References 34 publications

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“…As depicted in the transmission electron microscopy (TEM) images of Au-EJNC and Ag-EJNC, both nanoclusters exhibit fairly good water dispersion and ultrasmall size (Figure S4). The zeta potential also showed that both probes were negatively charged and had high absolute values (Figure S5), indicating that the dispersed particle system is stable. − We further measured the size of the nanoclusters by DLS. The size of the prepared Au and Ag nanoclusters is 3.0 and 2.4 nm, respectively (Figure S6).…”

Section: Resultsmentioning

confidence: 99%

Dual-Functional Nanocluster Probe-Based Single-Cell Analysis of RNA Splice Variants

Wang

Yang

et al. 2022

Anal. Chem.

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Differential expression of RNA splice variants among individual cells accounts for cell heterogeneity of gene expression, which plays a key role in the regulation of the immune system. However, currently available techniques face difficulties in achieving single-cell analysis of RNA splice variants with high base resolution, high spatial resolution and accurate quantification. Herein, we constructed DNA-templated dual-functional nanocluster probes to achieve in situ imaging and accurate quantification of RNA splice variants at the single-cell level. By designing ultrasmall nanocluster labeled probes to directly target the splicing junction sequence of RNA splice variants, the base recognition resolution is significantly improved. Benefit from the controllable fluorescence of nanoclusters, in situ imaging and genotyping of RNA splice variants are achieved. Due to the atom-precise nanocluster, RNA splice variants can be accurately quantified by laser ablation inductively coupled plasma mass spectrometry at the single-cell level. We further applied the probes to explore the function of MyD88 splice variants in mononuclear macrophages under immune activation. This strategy provides a novel single-cell analysis tool for studying the functional diversity of the immune system and splicing-related immune diseases.

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Section: Resultsmentioning

confidence: 99%

Dual-Functional Nanocluster Probe-Based Single-Cell Analysis of RNA Splice Variants

Wang

Yang

et al. 2022

Anal. Chem.

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“…In the presence of a catalyst and fluorescein-oacrylate (FA) monomer, the ATRP reaction was initiated to form polymer chains with multiple FAs. Nowadays, plenty of fluorescent nanomaterials have been successfully prepared to replace traditional organic dyes for protein kinase detection, including gold nanoclusters (AuNCs), QDs and graphene quantum dots (GQDs) [109,[119][120][121]. For instance, Wang et al developed a sensitive fluorescence assay for the activity of casein kinase II (CK2) based on the Zr 4+ ion-induced aggregation of phosphorylated peptide-GQDs [110].…”

Section: Fluorescence Biosensorsmentioning

confidence: 99%

Biosensors with Metal Ion–Phosphate Chelation Interaction for Molecular Recognition

Hao

Dong

et al. 2023

Molecules

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Biosensors show promising prospects in the assays of various targets due to their advantages of high sensitivity, good selectivity and rapid response. Molecular recognition is a key event of biosensors, which usually involves the interaction of antigen–antibody, aptamer–target, lectin–sugar, boronic acid–diol, metal chelation and DNA hybridization. Metal ions or complexes can specifically recognize phosphate groups in peptides or proteins, obviating the use of biorecognition elements. In this review, we summarized the design and applications of biosensors with metal ion–phosphate chelation interaction for molecular recognition. The sensing techniques include electrochemistry, fluorescence, colorimetry and so on.

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“…As one of the most important post-translational modifications, protein phosphorylation plays critical regulatory roles in cell cycle progression, proliferation, signal transduction, apoptosis, and immune responses. , However, aberrant kinase activity and protein phosphorylation can induce severe symptoms and diseases. , Therefore, protein kinases have become a class of therapeutic targets, and a number of protein kinase inhibitors have been applied in clinical trials for the treatment of different diseases including cancers, inflammatory and autoimmune diseases, and neurological disorders. − Thus, developing efficient analytical methods for evaluating protein kinase activity and the screening of its inhibitors is essential to the fundamental biochemical study, clinical diagnosis, and drug discovery. To date, various methods have been reported for monitoring protein kinase activity, including radiometric assay, colorimetry, , fluorescence, , , surface plasmon resonance, electrochemistry, , photoelectrochemistry, , electrochemical luminescence, quartz crystal microbalance, and mass spectrometry . However, these reported assays for protein kinase usually require the use of three separate components, that is, ATP, protein substrate, and the signaling probe.…”

Section: Introductionmentioning

confidence: 99%

All-in-One Luminescent Lanthanide Coordination Polymer Nanoprobe for Facile Detection of Protein Kinase Activity

et al. 2022

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This study developed a novel luminescent assay for kinase activity using metal–organic coordination polymer nanoparticles (Tb/ATP-Zn) as the probe. Tb/ATP-Zn, self-assembled by adenosine triphosphate (ATP), Zn2+, and Tb3+, is non-luminescent. Protein kinase A (PKA) can catalyze the transformation of ATP within Tb/ATP-Zn nanoparticles to adenosine diphosphate (ADP), which in turn effectively sensitizes the luminescence of Tb3+. Based on this mechanism, Tb/ATP-Zn can realize the facile luminescent “turn-on” sensing of protein kinase activity without the use of external ATP and substrate peptide. Under optimized conditions, the fluorescence intensities of Tb/ATP-Zn at 550 nm are linear with the PKA activity within a range of 0.3–1.5 U·μL–1. The LOD (S/N = 3) of this method is down to 0.001 U·μL–1. The presented assay also features high selectivity, long-term stability, fast response, and convenient operation. Furthermore, Tb/ATP-Zn was successfully employed for monitoring PKA activity in cell lysis solutions. Probe Tb/ATP-Zn is thus expectable to be a powerful tool for the practical study of PKA in relevant biological events.

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Colorimetric and electrochemical determination of the activity of protein kinase based on retarded particle growth due to binding of phosphorylated peptides to DNA – capped silver nanoclusters

Cited by 20 publications

References 34 publications

Dual-Functional Nanocluster Probe-Based Single-Cell Analysis of RNA Splice Variants

Dual-Functional Nanocluster Probe-Based Single-Cell Analysis of RNA Splice Variants

Biosensors with Metal Ion–Phosphate Chelation Interaction for Molecular Recognition

All-in-One Luminescent Lanthanide Coordination Polymer Nanoprobe for Facile Detection of Protein Kinase Activity

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