Fluorometric determination of the activity of inorganic pyrophosphatase and its inhibitors by exploiting the peroxidase mimicking properties of a two-dimensional metal organic framework

Mikrochim Acta. 2019 Feb 15;186(3):190. doi: 10.1007/s00604-019-3250-y.

Abstract

A copper(II)-based two-dimensional metal-organic framework with nanosheet structure (CuBDC NS) that possesses peroxidase (POx) mimicking activity was prepared. In the presence of hydrogen peroxide, the system catalyses the oxidation of terephthalic acid to a blue-fluorescent product (excitation = 315 nm; emission = 425 nm). Pyrophosphate has a very strong affinity for Cu2+ ion and blocks the POx-mimicking activity of the CuBDC NS. If, however, inorganic pyrophosphatase is present, the POx mimicking activity is gradually restored because pyrophosphate is hydrolyzed. The findings were used to design a method for the determination of the activity of inorganic pyrophosphatase by fluorometry. Fluorescence increases linearly in the 1-50 mU·mL-1 inorganic pyrophosphatase activity range. The limit of detection is 0.6 mU·mL-1 (S/N = 3). Graphical abstract A copper(II)-based two-dimensional metal-organic framework (CuBDC NS) is described that possesses POx-mimicking activity. Inorganic pyrophosphate (PPi) was hydrolyzed to phosphate in the presence of inorganic pyrophosphatase (PPase). Hence, it cannot coordinate with Cu2+ in CuBDC NS, its structure was well-conserved to catalyses the oxidation of terephthalic acid (H2BDC) to produce a blue fluorescent product (oxBDC) in the presence of hydrogen peroxide (H2O2).

Keywords: CuBDC nanosheet; Hydrogen peroxide; Inhibitors; Peroxidase mimicking activity; Two-dimensional metal-organic frameworks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomimetic Materials / chemistry*
  • Diphosphates / chemistry
  • Enzyme Assays / methods
  • Enzyme Inhibitors / analysis*
  • Fluorescence
  • Fluorometry / methods
  • Humans
  • Hydrogen Peroxide / chemistry
  • Inorganic Pyrophosphatase / blood*
  • Inorganic Pyrophosphatase / chemistry
  • Limit of Detection
  • Metal-Organic Frameworks / chemistry*
  • Peroxidase / chemistry
  • Phthalic Acids / chemistry

Substances

  • Diphosphates
  • Enzyme Inhibitors
  • Metal-Organic Frameworks
  • Phthalic Acids
  • diphosphoric acid
  • terephthalic acid
  • Hydrogen Peroxide
  • Peroxidase
  • Inorganic Pyrophosphatase