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  • A simple and visible colorimetric method through Zr(4+)-phosphate coordination for the assay of protein tyrosine phosphatase 1B and screening of its inhibitors.

A simple and visible colorimetric method through Zr(4+)-phosphate coordination for the assay of protein tyrosine phosphatase 1B and screening of its inhibitors.

The Analyst (2015-07-15)
Juan Zhang, Jun Lv, Xiaonan Wang, Defeng Li, Zhaoxia Wang, Genxi Li
ABSTRACT

Inhibitors of protein tyrosine phosphatase 1B (PTP1B) are promising agents for the treatment of type 2 diabetes and obesity, so a colorimetric method has been developed in this work for PTP1B assay and screening of its inhibitors. The method is based on the chelation effect of zirconium (Zr(4+)) ions on the phosphate group, which may induce aggregation of 4-aminophenylphosphate-functionalized gold nanoparticles (APP/AuNPs) and the corresponding color change of the testing solution. Owing to the dephosphorylation of PTP1B, the aggregation of AuNPs will be influenced by PTP1B since there is no coordination reactivity between Zr(4+) ions and 4-aminophenol, the hydrolyzed product of APP catalyzed by the enzyme. Therefore, a simple colorimetric method for the assay of PTP1B activity can be developed. Under the optimized experimental conditions, the ratios of absorbance at a wavelength of 650 nm to that at 522 nm vary linearly with the PTP1B activity in the range from 0.005 to 0.18 U mL(-1) with the lowest detection limit of 0.0017 U mL(-1). Moreover, using this proposed method, the inhibition effect of 6-chloro-3-formyl-7-methylchromone, betulinic acid, ursolic acid, and sodium orthovanadate on PTP1B activity can be tested with IC50 values of 10, 13, 9, and 1.1 μM, respectively. Therefore, this new method has great potential not only for the detection of PTP1B activity but also for the screening of the inhibitors.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium orthovanadate, ≥90% (titration)
SAFC
HEPES
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Protein Tyrosine Phosphatase 1B human, recombinant, expressed in E. coli
SAFC
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