title: A single electrochemical biosensor for detecting the activity and inhibition of both protein kinase and alkaline phosphatase based on phosphate ions induced deposition of redox precipitates.
abstract: Protein kinase (PKA) and alkaline phosphatase (ALP) are clinically relevant enzymes for a number of diseases. In this work, we developed a new simple electrochemical biosensor for the detection of the activity and inhibition of both PKA and ALP. One common feature of the PKA and ALP catalyzing process is that PKA can hydrolysis adenosine-5'-triphosphate (ATP) and ALP can hydrolysis pyrophosphate, both reactions produce phosphate ions, and the amount of phosphate ion produced is proportional to enzyme activity. Our assay is based on the principle that phosphate ions react with molybdate to form redox molybdophosphate precipitates on the electrode surface, thus generating electrochemical current. The detection limit for PKA and ALP were much lower than existing assays. The biosensor has good specificity and was used to measure drug-stimulated PKA from lysates of HeLa cells. We also evaluated the use of the biosensor as a screening tool for enzyme inhibitors. To the best of our knowledge, this is the first report of a biosensor capable of detecting the activity of both PKA and ALP. This tool has the potential to simplify PKA and ALP clinical measurement, thereby improving diagnostics of relevant diseases. It also may serve as the basis for a simple screening method for new enzyme inhibitors for disease treatment.
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Congcong
authors.initials: C
authors.lastname: Shen
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Xiangzhi
authors.initials: X
authors.lastname: Li
authors.affiliation: National Cancer Institute, National Institutes of Health, Rockville, MD 20850, USA.
authors.forename: Avraham
authors.initials: A
authors.lastname: Rasooly
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Linyan
authors.initials: L
authors.lastname: Guo
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Kaina
authors.initials: K
authors.lastname: Zhang
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. Electronic address: yangminghui@csu.edu.cn.
authors.forename: Minghui
authors.initials: M
authors.lastname: Yang
mesh_terms: D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D055664:Electrochemical Techniques / Q000379:methods; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008982:Molybdenum / Q000378:metabolism; D010084:Oxidation-Reduction; D010710:Phosphates / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D010710:Phosphates; D047428:Protein Kinase Inhibitors; C044659:molybdate; D008982:Molybdenum; D011494:Protein Kinases; D000469:Alkaline Phosphatase
keywords: Alkaline phosphatase; Biosensor; Electrochemical; Inhibitor; Protein kinase