We developed a novel enzyme cycling method using hypoxanthine-guanine phosphoribosyltransferase (HGPRT) (EC 2.4.2.8) from . This method exploits the reversible nature of the HGPRT reaction, catalyzing both forward and reverse reactions in the presence of excess inosine 5'-monophosphate (IMP) and guanine (Gua), similar to the established purine nucleoside phosphorylase (PNP)-based method. Real-time detection was achieved by coupling the reaction with commercially available xanthine dehydrogenase (XDH) (EC 1.17.1.4) in the presence of nicotinamide adenine dinucleotide (NAD). The reaction exhibited high efficiency, with a cycling rate constant of approximately 60 min or higher at an HGPRT concentration of 1 U mL. Additionally, we demonstrated a preliminary application of this XDH-coupled enzyme cycling reaction for the determination of pyrophosphate (PPi).
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1039/d4ay01692k | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Enzyme cycling method using hypoxanthine-guanine phosphoribosyltransferase: a highly sensitive assay for pyrophosphate.
Anal Methods
March 2025
R&D Group, Diagnostics Dept., Asahi Kasei Pharma Corporation, Izunokuni 410-2321, Japan.
We developed a novel enzyme cycling method using hypoxanthine-guanine phosphoribosyltransferase (HGPRT) (EC 2.4.2.
View Article and Find Full Text PDFThe role of mA modification during macrophage metabolic reprogramming in human diseases and animal models.
Front Immunol
March 2025
Department of Laboratory Medicine, Jiangsu Province Engineering Research Center for Precise Diagnosis and Treatment of Inflammatory Diseases, The Affiliated Hospital of Jiangsu University, Zhenjiang, China.
Macrophage metabolic reprogramming refers to the process by which macrophages adjust their physiological pathways to meet survival and functional demands in different immune microenvironments. This involves a range of metabolic pathways, including glycolysis, the tricarboxylic acid cycle, oxidative phosphorylation, fatty acid oxidation, and cholesterol transport. By modulating the expression and activity of key enzymes and molecules within these pathways, macrophages can make the transition between pro- and anti-inflammatory phenotypes, thereby linking metabolic reprogramming to inflammatory responses and the progression of several diseases, such as atherosclerosis, inflammatory bowel disease (IBD), and acute lung injury (ALI).
View Article and Find Full Text PDFNew Insight Into the Mechanism of Nitrite Enhancement on Heterotrophic Nitrification and Aerobic Denitrification Bacterium in Gene Expression.
Environ Microbiol
March 2025
Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences, Guizhou University, Guiyang, Guizhou, China.
The growth and nitrogen metabolism of heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria are affected by nitrite, but the mechanisms underlying this for strain Acinetobacter johnsonii EN-J1 are unclear. In this study, the addition of 10 mg/L nitrite increased the reduction rate of ammonium by 1.0 mg/L/h, and 20 mg/L nitrite increased the reduction rate of nitrate by 3.
View Article and Find Full Text PDFAccelerating discovery of bioactive ligands with pharmacophore-informed generative models.
Nat Commun
March 2025
Center for Quantitative Biology, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China.
Deep generative models have advanced drug discovery but often generate compounds with limited structural novelty, providing constrained inspiration for medicinal chemists. To address this, we develop TransPharmer, a generative model that integrates ligand-based interpretable pharmacophore fingerprints with a generative pre-training transformer (GPT)-based framework for de novo molecule generation. TransPharmer excels in unconditioned distribution learning, de novo generation, and scaffold elaboration under pharmacophoric constraints.
View Article and Find Full Text PDFAssessment of L-cysteine Reductase Activity as an Indicator of Soil Health in Different Climatic Zones After Pollution by Oil.
Bull Environ Contam Toxicol
March 2025
Academy of Biology and Biotechnology by D.I. Ivanovsky, Southern Federal University, Rostov-on-Don, 344090, Russia.
The sulfur cycle in the soil regulates many vital processes in plants since it enters the composition of amino acids, vitamins, phytochelatins, and other substances. Soils of different climatic zones have different physicochemical properties and sulfur reserves, which, therefore, stipulates various activities of soil enzymes. The aim of the study was the assessment of L-cysteine reductase activity as an indicator of soil health in different climatic zones after oil pollution: semi-desert types, steppe types and forest types of soils.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!