A monomer-dimer switch modulates the activity of plant adenosine kinase

Adenosine undergoes ATP-dependent phosphorylation catalyzed by adenosine kinase (ADK). In plants, ADK also phosphorylates cytokinin ribosides, transport forms of the hormone. Here, we investigated the substrate preferences, oligomeric states and structures of ADKs from moss (Physcomitrella patens) and maize (Zea mays) alongside metabolomic and phenotypic analyses. We showed that dexamethasone-inducible ZmADK overexpressor lines in Arabidopsis can benefit from a higher number of lateral roots and larger root areas under nitrogen starvation. We discovered that maize and moss enzymes can form dimers upon increasing protein concentration, setting them apart from the monomeric human and protozoal ADKs. Structural and kinetic analyses revealed a catalytically inactive unique dimer. Within the dimer, both active sites are mutually blocked. The activity of moss ADKs, exhibiting a higher propensity to dimerize, was tenfold lower compared to maize ADKs. Two monomeric structures in a ternary compl..., This collection of datasets is related to molecular properties, ligand interactions and enzyme kinetics. Data measurements are given in Materials and methods. Other data are part of the Supplement of the manuscript.
Gel permeation chromatography
Gel permeation chromatography of studied plant Aadenosine kinases was performed on an NGC Medium-Pressure Liquid Chromatography System (https://www.bio-rad.com)on a Superdex 200 10/30 HR column in 20 mM Tris-HCl buffer, pH 7.5, 100 mM NaCl, with calibration performed using a gel filtration standard (Bio-Rad).
Affinity and thermal stability measurements
The MST method was used to determine the binding affinity of various ribosides to ZmADK2 and PpADK1. Proteins were fluorescently labeled with RED-tris-NTA dye (www.nanotemper-technologies.com) using a 1:1 dye/protein molar ratio. The labeled protein was adjusted to 100-300 nM in 50 mM HEPES buffer pH 7.5, 1 mM MgCl2 and 0.2% Tween. Measurements were performed in premium capillaries on a Monolith N..., , # Data from: A monomer-dimer switch modulates the activity of plant adenosine kinase

[https://doi.org/10.5061/dryad.qrfj6q5sj](https://doi.org/10.5061/dryad.qrfj6q5sj)

## Description of the data and file structure

This supplementary dataset, integral to our research paper, contains data covering molecular properties and interactions of studied plant adenosine kinases. It includes final microscale thermophoresis data used to describe the binding curves and, thus, affinity to various riboside ligands. The dataset consists of gel permeation chromatography profiles of studied plant adenosine kinases as well as the dynamic light scattering (DLS) data used to determine the oligomeric state of studied enzymes. Thermal stability curves measured by nano-differential scanning fluorimetry are provided to show differences in stability in the presence of ATP. The dataset further contains kinetic data measured with maize ADK2 used to determine Km and Vmax values. Finally, quantitative PCR (qPCR) d...,