Thiophosphate linkages are increasingly valued as metabolic-stability-enhancing replacements for natural phosphate groups, enabling longer-lived chemistry in therapeutic contexts. Their use spans antisense oligonucleotides and cyclic dinucleotide analogs, where resistance to enzymatic breakdown can translate into improved biological performance. Yet installing thiophosphates biocatalytically has remained a bottleneck.
The conventional enzymatic approach relies on protein kinases using adenosine-5′-O-(3-thio-triphosphate) (ATPγS) as the sulfur donor. While effective, this strategy has two practical drawbacks: it has largely been confined to limited substrate types, and it is difficult to scale because ATPγS is costly and typically required in superstoichiometric amounts. In most kinase cycles, the reaction consumes ATPγS and produces a kinase byproduct that is not readily reused.
In a new study appearing in Nature, Wu, Fu, and Renata report an ATPγS recycling concept designed to make kinase-based thiophosphorylation operationally practical. The method uses a tailored creatine-derived component that is able to turnover the kinase reaction byproduct, effectively restoring the productive ATPγS pool. As a result, the overall process can maintain thiophosphate installation while reducing the need for expensive ATPγS inputs.
Crucially, the recycling architecture is not tied to a single kinase. The authors describe compatibility with many kinase families, broadening the substrate and enzyme space accessible to thiophosphate synthesis. This widens the scope beyond what earlier ATPγS-based methods could reliably achieve.
To demonstrate versatility, the team builds multi-enzyme cascades that couple kinase activity with ATPγS regeneration and downstream transformations. These cascades enable access to multiple thiophosphate-containing targets in one operational framework, rather than requiring separate, discontinuous reaction setups.
Among the products are nucleoside 5′-monothiophosphates, 3′,5′-cyclic monophosphorothioates, and thiophosphorylated oligopeptides. The ability to generate both small-molecule and macromolecular thiophosphates highlights the method’s generality and potential utility in drug-development synthesis.
For viral science news readers, the broader implication is clear: thiophosphates can modulate stability and recognition, properties central to nucleic-acid and signaling therapeutics that may intersect with antiviral development pipelines. By lowering the cost and complexity of thiophosphate installation, this enzymatic platform could accelerate discovery and optimization of thiophosphate-containing candidates.
Overall, the reported ATPγS recycling strategy reframes thiophosphorylation from a specialized, ATPγS-intensive technique into a more scalable biocatalytic workflow, opening a path to new synthesis routes for next-generation phosphate-mimic modalities.
Subject of Research: ATPγS recycling strategy for biocatalytic thiophosphorylation
Article Title: An ATPγS recycling strategy for practical biocatalytic thiophosphorylation
Article References: Wu, X., Fu, Y. & Renata, H. An ATPγS recycling strategy for practical biocatalytic thiophosphorylation. Nature (2026). https://doi.org/10.1038/s41586-026-10895-9
Image Credits: AI Generated
DOI: 10.1038/s41586-026-10895-9
Keywords: thiophosphate; ATPγS; protein kinases; biocatalysis; enzyme cascades; creatine derivative; phosphorothioate; metabolic stability
Tags: ATPγS recyclingbiocatalytic thiophosphorylationbroad kinase compatibility in thiophosphorylationcost-effective methods for kinase-catalyzed modificationscreatine-derived ATPγS regenerationenzymatic synthesis of thiophosphate-modified biomoleculesenzyme engineering for phosphate analogskinase reaction optimizationmetabolic stability of nucleic acidsscalable biocatalytic phosphorylation methodssulfur donor reuse in enzymatic reactionsthiophosphate linkages in therapeutics
