Electrochemical detection of anti-tau antibodies binding to tau protein and inhibition of GSK-3β-catalyzed phosphorylation
| Autor: | Sanela Martic-Milne, Jose O. Esteves-Villanueva |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Surface Properties Tau protein Biophysics Spectrometry Mass Secondary Ion Hyperphosphorylation Phosphorylation Inhibition tau Proteins Biochemistry Antibodies Epitope Glycogen Synthase Kinase 3 03 medical and health sciences Phosphorylation Protein kinase A Molecular Biology Glycogen Synthase Kinase 3 beta biology Kinase Chemistry Photoelectron Spectroscopy Cell Biology 030104 developmental biology Toxicity Biocatalysis biology.protein |
| Zdroj: | Analytical Biochemistry. 496:55-62 |
| ISSN: | 0003-2697 |
| Popis: | Tau protein hyperphosphorylation triggers tau aggregation and its toxicity, leading to neuronal death and cell-to-cell toxicity. Hence, inhibition of protein kinases is a viable tool toward reduction of tau toxicity. By targeting various epitopes of Tau441 protein immobilized on Au surface, the protein kinase inhibition by anti-tau antibodies was measured by surface electrochemistry. The electrochemical impedance spectroscopy was used to measure the charge transfer resistance (Rct) of nonphosphorylated tau–Au film (nTau–Au) and compared with the phosphorylated tau–Au film (pTau–Au). The pTau–Au films were characterized by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (TOF–SIMS), which indicated high phosphorus content. The Rct factor was used as the measure of inhibition efficacies by anti-tau antibodies (D8, A10, P262, and Tau46) in addition to antibody formulation intravenous immunoglobulin (IVIG). The Rct factor for pTau–Au in the absence of antibodies was 0.25 ± 0.08, indicating a dramatic decrease in Rct on phosphorylation. The Rct factors for Tau46 and A10 were 0.57 ± 0.22 and 0.65 ± 0.26, respectively, indicating phosphorylation inhibition. All antibodies exhibited similar binding to nTau–Au. The proposed electrochemical assay may be used for detection of other posttranslational modifications. |
| Databáze: | OpenAIRE |
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