Electrochemical biosensor for protein kinase A activity assay based on gold nanoparticles-carbon nanospheres, phos-tag-biotin and β-galactosidase

2016 ◽  
Vol 86 ◽  
pp. 508-515 ◽  
Author(s):  
Yunlei Zhou ◽  
Huanshun Yin ◽  
Xue Li ◽  
Zhi Li ◽  
Shiyun Ai ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Electrochemical Biosensor ◽  
Activity Assay ◽  
Carbon Nanospheres ◽  
Kinase A

scholarly journals Phosphorylation-Dependent SERS Readout for Activity Assay of Protein Kinase A in Cell Extracts

Nanomaterials ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 575
Author(s):  
Renyong Liu ◽  
Chenggen Xie ◽  
Yehan Yan ◽  
Lin Hu ◽  
Suhua Wang ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Biological Samples ◽  
Cell Function ◽  
Activity Assay ◽  
Cell Extracts ◽  
Disease Diagnostics ◽  
Wide Range ◽  
Surface Enhanced Raman ◽  
Kinase A

Protein kinases are key regulators of cell function, the abnormal activity of which may induce several human diseases, including cancers. Therefore, it is of great significance to develop a sensitive and reliable method for assaying protein kinase activities in real biological samples. Here, we report the phosphorylation-dependent surface-enhanced Raman scattering (SERS) readout of spermine-functionalized silver nanoparticles (AgNPs) for protein kinase A (PKA) activity assay in cell extracts. In this assay, the presence of PKA would phosphorylate and alter the net charge states of Raman dye-labeled substrate peptides, and the resulting anionic products could absorb onto the AgNPs with cationic surface charge through electrostatic attraction. Meanwhile, the Raman signals of dyes labeled on peptides were strongly enhanced by the aggregated AgNPs with interparticle hot spots formed in assay buffer. The SERS readout was directly proportional to the PKA activity in a wide range of 0.0001–0.5 U·μL−1 with a detection limit as low as 0.00003 U·μL−1. Moreover, the proposed SERS-based assay for the PKA activity was successfully applied to monitoring the activity and inhibition of PKA in real biological samples, particularly in cell extracts, which would be beneficial for kinase-related disease diagnostics and inhibitor screening.

Ultrasensitive amplification-free detection of protein kinase based on catalyzed assembly and enumeration of gold nanoparticles

2019 ◽  
Vol 55 (17) ◽  
pp. 2505-2508 ◽  
Author(s):  
Tongtong Tian ◽  
Yuanyuan Yao ◽  
Beibei Yang ◽  
Kun Zhang ◽  
Baohong Liu
Keyword(s):  
Gold Nanoparticles ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Single Particle ◽  
Label Free ◽  
Kinase A

A novel single-particle enumeration method for label-free, amplification-free and ultrasensitive probing of protein kinase A activity based on catalyzed assembly.

Facile and Sensitive Method for Protein Kinase A Activity Assay Based on Fluorescent Off-On PolyU-peptide Assembly

2017 ◽  
Vol 89 (20) ◽  
pp. 10980-10984 ◽  
Author(s):  
Yanhui Xu ◽  
Wen Shi ◽  
Xinyuan He ◽  
Xiaofeng Wu ◽  
Xiaohua Li ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Activity Assay ◽  
Peptide Assembly ◽  
Sensitive Method ◽  
Kinase A

A peptide array-based serological protein kinase A activity assay and its application in cancer diagnosis

The Analyst ◽  
2015 ◽  
Vol 140 (19) ◽  
pp. 6588-6594 ◽  
Author(s):  
Deok-Hoon Kong ◽  
Se-Hui Jung ◽  
Hye-Yoon Jeon ◽  
Woo-Jin Kim ◽  
Young-Myeong Kim ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Sensitivity And Specificity ◽  
Cancer Diagnosis ◽  
Crucial Role ◽  
Peptide Array ◽  
Biological Processes ◽  
Activity Assay ◽  
P Protein ◽  
Kinase A

Protein kinase A (PKA) plays a crucial role in several biological processes; however, there is no assay with sufficient sensitivity and specificity to determine serological PKA (sPKA) activity.

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