Measurement of Homogeneous Kinase Activity for Cell Lysates Based on the Aggregation of Gold Nanoparticles

Oishi, Jun; Asami, Yoji; Mori, Takeshi; Kang, Jeong Hun; Tanabe, Miharu; Niidome, Takuro; Katayama, Yoshiki

doi:10.1002/cbic.200700086

Cited by 75 publications

(52 citation statements)

References 27 publications

(8 reference statements)

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“…However, this method is rapid (,30 min), does not require a phosphorylation process with the use of derived ATP cosubstrate, and obviates the modification of nanoparticles for signal amplification. Because the charge of kinase-specific peptide probe (inhibitor) could exactly be tuned by inserting positively or negatively charged amino acid residues and AuNPs in the both positive and negative forms can be easily prepared, 12,53,54,69 it is believed that the present …”

Section: Resultsmentioning

confidence: 99%

“…Usually, there are two categories of the colorimetric sensors based on the AuNPs used: one is based on the cross-linking aggregation of AuNPs functionalized respectively with recognition elements (eg, avidin and antibody) and kinase-specific peptide, 10,11,14,15 and the other is based on the non-cross-linking aggregation of unmodified AuNPs, which is tuned by the phosphorylation-induced net charge change of substrate peptide. 12,53,54 Although the cross-linking strategies are robust and highly specific, they suffer from the complicated functionalization of AuNPs. The unmodified method is simple and does not require modification of analyte-binding molecules onto the surface of AuNPs, but it shows poor anti-interference ability to high concentration of salts and other components in real samples.…”

mentioning

confidence: 99%

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Gold nanoparticles-based electrochemical method for the detection of protein kinase with a peptide-like inhibitor as the bioreceptor

Sun¹,

Chang²,

Zhou³

et al. 2017

IJN

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This article presents a general method for the detection of protein kinase with a peptide-like kinase inhibitor as the bioreceptor, and it was done by converting gold nanoparticles (AuNPs)-based colorimetric assay into sensitive electrochemical analysis. In the colorimetric assay, the kinase-specific aptameric peptide triggered the aggregation of AuNPs in solution. However, the specific binding of peptide to the target protein (kinase) inhibited its ability to trigger the assembly of AuNPs. In the electrochemical analysis, peptides immobilized on a gold electrode and presented as solution triggered together the in situ formation of AuNPs-based network architecture on the electrode surface. Nevertheless, the formation of peptide–kinase complex on the electrode surface made the peptide-triggered AuNPs assembly difficult. Electrochemical impedance spectroscopy was used to measure the change in surface property in the binding events. When a ferrocene-labeled peptide (Fc-peptide) was used in this design, the network of AuNPs/Fc-peptide produced a good voltammetric signal. The competitive assay allowed for the detection of protein kinase A with a detection limit of 20 mU/mL. This work should be valuable for designing novel optical or electronic biosensors and likely lead to many detection applications.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Gold nanoparticles-based electrochemical method for the detection of protein kinase with a peptide-like inhibitor as the bioreceptor

Sun¹,

Chang²,

Zhou³

et al. 2017

IJN

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“…Different methods for the detection of protein kinases have been developed, such as immunoassays that use fluorescence-labeled antibodies (Fab) against sequence-specific phosphorylated peptides, [4] and the protein kinase-induced radioactive labeling of the phosphorylated product with radioactive ATP. [5] Other methods to monitor the activities of protein kinases include the fluorescence polarization assay, [6] the amplified electrochemical detection of kinase with Au nanoparticles, [7] a protein kinase-induced aggregation of Au nanoparticles, [8] and detection of the depletion of ATP as a result of a biocatalyzed phosphorylation process. [9] Furthermore, a label-free detection of protein kinase was achieved by monitoring the phosphorylation reaction on a field-effect transistor device.…”

Section: Introductionmentioning

confidence: 99%

Probing Kinase Activities by Electrochemistry, Contact‐Angle Measurements, and Molecular‐Force Interactions

Wilner

Guidotti

Więckowska

et al. 2008

Chemistry A European J

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Three different methods to investigate the activity of a protein kinase (casein kinase, CK2) are described. The phosphorylation of the sequence-specific peptide (1) by CK2 was monitored by electrochemical impedance spectroscopy (EIS). Phosphorylation of the peptide monolayer assembled on a Au electrode yields a negatively charged surface that electrostatically repels the negatively charged redox label [Fe(CN)6]3-/4-, thus increasing the interfacial electron-transfer resistance. The phosphorylation process by CK2 is further amplified by the association of the anti-phosphorylated peptide antibody to the monolayer. Binding of the antibody insulates the electrode surface, thus increasing the interfacial electron-transfer resistance in the presence of the redox label. This method enabled the quantitative analysis of the concentration of CK2 with a detection limit of ten units. The second method employed involved contact-angle measurements. Although the peptide 1-functionalized electrode revealed a contact angle of 67.5 degrees , phosphorylation of the peptide yielded a surface with enhanced hydrophilicity, 36.8 degrees. The biocatalyzed cleavage of the phosphate units with alkaline phosphatase regenerates the hydrophobic peptide monolayer, contact angle 55.3 degrees . The third method to characterize the CK2 system involved chemical force measurements between the phosphorylated peptide monolayer associated with the Au surface and a Au tip functionalized with the anti-phosphorylated peptide antibody. Although no significant rupture forces existed between the modified tip and the 1-functionalized surface (6+/-2 pN), significant rupture forces (multiples of 120+/-20 pN) were observed between the phosphorylated monolayer-modified surface and the antibody-functionalized tip. This rupture force is attributed to the dissociation of a simple binding event between the phosphorylated peptide and the fluorescent antibody (Fab) binding region.

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“…2). [18] In addition to monitoring the activity of purified enzymes, this simple colorimetric approach could also be extended to observing the effects of stimulatory drugs on cellular signal-transduction pathways that enhance kinase activity. An analogous procedure has also been developed to detect the activity of phosphatases; enzymes that catalyze the removal of phosphate groups from proteins.…”

Section: Protein Kinasesmentioning

confidence: 99%

“…a) Kinasemediated crosslinking of gold nanoparticles by means of a biotinylated substrate analogue [17]. b) Utilization of changes in peptide charge following phosphorylation to influence nanoparticle aggregation [18].…”

Section: Protein Kinasesmentioning

confidence: 99%

Enzyme‐Responsive Nanoparticle Systems

2008

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Inorganic nanoparticles and their accompanying diverse physical properties are now virtually in routine use as imaging tools in cell‐biology. In addition to serving as excellent contrast agents, their size‐ and environment‐dependent optical and magnetic properties can be harnessed to create enzyme biosensor devices of extremely high sensitivity, whilst circumventing the numerous technical limitations associated with traditional enzyme assays. In this Research News article we discuss recent advances in field of enzyme‐responsive nanoparticle systems, where the activity of an enzyme elicits a specific response in the nanoparticle assembly to produce a signal relating to enzyme activity, focusing on three important systems: DNA‐structured nanoparticles, protein kinases and proteases.

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Measurement of Homogeneous Kinase Activity for Cell Lysates Based on the Aggregation of Gold Nanoparticles

Cited by 75 publications

References 27 publications

Gold nanoparticles-based electrochemical method for the detection of protein kinase with a peptide-like inhibitor as the bioreceptor

Gold nanoparticles-based electrochemical method for the detection of protein kinase with a peptide-like inhibitor as the bioreceptor

Probing Kinase Activities by Electrochemistry, Contact‐Angle Measurements, and Molecular‐Force Interactions

Enzyme‐Responsive Nanoparticle Systems

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