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Zr4+-mediated hybrid chain reaction and its application for highly sensitive electrochemical detection of protein kinase A.
Cheng, Wenting; Ma, Jiehua; Xiang, Liangliang; Sun, Ying; Huang, Wei; Zhang, Zhaoli; Kong, Dehua; Li, Jinlong.
Afiliación
  • Cheng W; Department of Clinical Laboratory, Nanjing Gaochun People's Hospital, Nanjing 211300, PR China.
  • Ma J; Women's Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing 210004, PR China.
  • Xiang L; The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210003, PR China.
  • Sun Y; Department of Clinical Laboratory, Nanjing Gaochun People's Hospital, Nanjing 211300, PR China.
  • Huang W; Department of Clinical Laboratory, Nanjing Gaochun People's Hospital, Nanjing 211300, PR China.
  • Zhang Z; The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210003, PR China.
  • Kong D; Department of Clinical Laboratory, Nanjing Gaochun People's Hospital, Nanjing 211300, PR China. Electronic address: 13851912568@163.com.
  • Li J; The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210003, PR China. Electronic address: jinlonglilab@126.com.
Bioelectrochemistry ; 140: 107796, 2021 Aug.
Article en En | MEDLINE | ID: mdl-33744680
An electrochemical platform has been developed to detect protein kinase activity through the combined actions of Zr4+ mediated signal transition and hybridization chain reaction (HCR)-stimulated DNAzymes nanowires. First of all, protein kinase A (PKA) phosphorylates substrate peptides immobilized on gold electrode surface. Thereafter, the DNA1 containing 5'-phosphoryl ends is linked to the phosphorylated substrate peptide via the robust phosphate-Zr4+-phosphate linkages. By the introduction of molecular beacons (MBs), the DNA1 can open the hairpin structures of MBs through toehold mediated strand displacement (TMSDR), leading to an autonomous stem-opening process and subsequent assembly of G-quadruplex-containing DNA chains by HCR. After the addition of hemin, the formed HRP-mimicking DNAzymes can catalyze the hydroquinone-H2O2 system to generate amplified electrochemical signals. As expected, this method can achieve ultrahigh analytical performance with a low detection limit of 0.02U/mL and exhibit high cost-savings potential without the need for antibody, protease and labeling. Therefore, this method can serve as a new tool for the assay of protein kinase A and its inhibitor screening in the future.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Circonio / Proteínas Quinasas Dependientes de AMP Cíclico / Pruebas de Enzimas Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Bioelectrochemistry Asunto de la revista: BIOQUIMICA Año: 2021 Tipo del documento: Article Pais de publicación: Países Bajos
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Circonio / Proteínas Quinasas Dependientes de AMP Cíclico / Pruebas de Enzimas Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Bioelectrochemistry Asunto de la revista: BIOQUIMICA Año: 2021 Tipo del documento: Article Pais de publicación: Países Bajos