Single-strand DNA-scaffolded copper nanoclusters for the determination of inorganic pyrophosphatase activity and screeni
- PDF / 1,754,427 Bytes
- 8 Pages / 595.276 x 790.866 pts Page_size
- 0 Downloads / 169 Views
ORIGINAL PAPER
Single-strand DNA-scaffolded copper nanoclusters for the determination of inorganic pyrophosphatase activity and screening of its inhibitor Jiawei Pang 1 & Yuexiang Lu 2 & Xinyu Gao 1 & Liuying He 1 & Jingwei Sun 1 & Fengyi Yang 1 & Yueying Liu 1 Received: 14 November 2019 / Accepted: 15 November 2020 / Published online: 23 November 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract A fluorescence method for the determination of inorganic pyrophosphatase (PPase) activity has been established based on copper nanoclusters (CuNCs). The polythymine of 40 mer (T40) acts as a template for the reduction reaction from Cu2+ to Cu0 by ascorbic acid (AA). This reaction leads to the formation of fluorescent CuNCs with excitation/emission peaks at 340/640 nm. However, the higher binding affinity between inorganic pyrophosphate (PPi) and Cu2+ hinders the effective formation of CuNCs. This shows low fluorescence intensity. PPase catalyzes the hydrolysis of PPi into Pi during which free Cu2+ ions are produced. This facilitates the formation of fluorescent CuNCs. Thus, the fluorescence intensity was restored. The fluorescence enhancement of the system has a linear relationship with PPase activity in the range 0.3 to 20 mU·mL−1, and the detection limit is0.2 mU·mL−1. The relative intensity (I/I0) at 640 nm for the analytical solution versus system is also employed to screen the inhibitor for PPase with high efficiency. Keywords Polythymine template . Copper nanoclusters . PPase activity assay . Enzyme inhibitor . Pyrophosphate . Single-strand DNA . Fluorescent probe . PolyT40scaffold
Introduction Pyrophosphatase (EC 3.6.1.1), or inorganic pyrophosphatase (PPase), can specifically catalyze the hydrolysis of one molecule of inorganic pyrophosphate (P2O74−, PPi) into two molecules of orthophosphate (Pi). This decomposition reaction provides a thermodynamic pull for biosynthetic reactions [1]. Some studies have revealed that PPase plays a crucial role in many biological processes, such as phosphorus and carbohydrate metabolism, lipid metabolism, calcium absorption, and other biochemical transformations [2]. In addition, overexpression and aberrant activity of PPase may be an indication or a cause of multiple complex diseases, including colorectal
* Yueying Liu [email protected] 1
Department of Chemistry, Capital Normal University, Xisanhuan North Rd. 105, Beijing 100048, People’s Republic of China
2
Institute of Nuclear and New Energy Technology, Collaborative Innovation Center of Advanced Nuclear Energy Technology, Beijing Key Lab of Radioactive Waste Treatment, Tsinghua University, Beijing 100084, People’s Republic of China
cancer, hyperthyroidism, and lung adenocarcinomas [3]. Therefore, the development of sensitive PPase detection methods is of great significance to understanding its biological functionality and exploring its application in disease diagnosis. To date, most efforts have been devoted to developing several methods for the detection of PPase activity, including radioact
Data Loading...
