Adsorption between TC-stabilized AuNPs and the phosphate group: application of the PTP1B activity assay

Abstract

Based on the adsorption between tetracycline (TC) and phosphate groups, a general colorimetric method is explored in this work by using TC-stabilized gold nanoparticles (TC/AuNPs) and 4-aminophenyl phosphate-functionalized Fe₃O₄ magnetic nanoparticles (APP/MNPs). Taking protein tyrosine phosphatase 1B (PTP1B) as an example, 4-aminophenyl phosphate (APP) can be hydrolyzed into 4-aminophenol (AP) by PTP1B, resulting in the disappearance of the phosphate group on the outer layer of MNPs and the loss of corresponding adsorptive ability. Upon addition of TC/AuNP solution, TC/AuNPs will remain in the supernatant solution after magnetic separation and a high absorbance value can be observed. So PTP1B activity is related to the concentrations of TC/AuNPs in the supernatant solution. In this work, the enzyme activity can be determined at levels as low as 0.0885 U mL⁻¹ and over a linear detection range as wide as 0.1 U mL⁻¹ to 0.9 U mL⁻¹. Moreover, using the proposed method, the inhibition effect of betulinic acid (BA) and sodium orthovanadate (Na₃VO₄) on PTP1B activity can be tested with IC₅₀ values of 30 μM and 4 μM, respectively. Therefore, a universal platform for the accurate colorimetric analysis of kinase and phosphatase activities can be established through the adsorption between TC and phosphate groups.