Biology subjects

Kopecny, D.

Publications and source records attributed to Kopecny, D..

6 recordsLinked to original sources

bioRxiv2026

Structural basis for far-red light harvesting in a euglenophyte photosystem II supercomplex

Photosystem II (PSII) is in eukaryotic phototrophs is generally considered to operate within a more restricted spectral range than photosystem I (PSI), in which long-wavelength chlorophylls are a well-established feature of the peripheral antenna. Whether eukaryotic PSII can acquire comparable far-red-associated properties through lineage-specific antenna diversification has remained unclear. Here we present a 3.09 [A] cryo-electron microscopy structure of the C2S2M2L2 PSII supercomplex from Euglena gracilis, a euglenophyte species harbouring a secondary plastid and unusual light-harvesting system. We show that the euglenophyte-specific antenna protein LhcE9 occupies the position corresponding to canonical Lhcb5, but in a markedly different orientation that creates a distinct interface with the PSII core, particularly with CP43. Combined structural, spectroscopic, mutagenesis and proteomic analyses support LhcE9 as the stably bound PSII antenna subunit most closely associated with the far-red state in the supercomplex. Excitation-energy-transfer calculations further indicate two fast lineage-specific antenna-to-core routes mediated by LhcE9 and PsbX. Together, these findings reveal an unexpected mode of PSII antenna diversification and provide a structural framework for far-red-associated light harvesting in PSII.

plant biology↗

bioRxiv2026

Insight into GABA shunt-associated aldehyde dehydrogenases (ALDH) and stress responses of ALDH superfamily in moss and barley

We explored the expression of the aldehyde dehydrogenase (ALDH) superfamily in two model plants, Physcomitrium patens (moss) and Hordeum vulgare (barley), under various stress conditions. The ALDH enzymes are crucial for oxidizing aldehydes to carboxylic acids and are involved in multiple metabolic pathways. We found significant differences in enzyme expression between moss and barley within the same ALDH families. We then focused on the ALDH5, ALDH10, and ALDH21 families, which are part of the {gamma}-aminobutyric acid (GABA) shunt, noting that the ALDH21 family is absent in barley. The kinetic properties of ALDH10 and ALDH5 enzymes were analyzed, revealing that PpALDH5F1 exhibits high specificity for succinic semialdehyde (SSAL), a product of GABA. The crystal structure of PpALDH5F1 identified key residues for SSAL binding. Knockout mutants of moss aldh5F2, aldh10A1, and aldh21A1 showed slightly smaller colonies than the wild-type. GABA and glutamate levels were elevated in aldh5F2 and aldh21A1 knockouts due to a partially blocked GABA shunt pathway, while aldh10A1 knockout showed no changes in GABA levels. Transcriptomic data revealed a link between several genes, including six upregulated glutathione-S-transferase genes in all three aldh knockouts, suggesting a direct compensatory mechanism for oxidative stress protection via conjugation of undegraded aldehydes to glutathione. HighlightMoss knockouts of GABA shunt-associated aldehyde dehydrogenases display slower growth, changes in levels of glutamate, glutamine and GABA, and result in upregulation of several unique glutathione-S-transferase genes.

plant biology↗

bioRxiv2024

The monomer/dimer switch modulates the activity of plant adenosine kinase

HighlightThe switch from active monomers to inactive dimers in plant ADKs impacts overall enzyme activity and represents a novel negative feedback-loop mechanism to maintain steady levels of adenosine and AMP. 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 complex highlight the characteristic transition from an open to a closed state upon substrate binding. This suggests that the oligomeric state switch can modulate the activity of moss ADKs and likely other plant ADKs. Moreover, dimer association represents a novel negative feedback mechanism, helping to maintain steady levels of adenosine and AMP.

plant biology↗

bioRxiv2024

Phenylacetic acid metabolism in land plants: novel pathways and metabolites

In recent years, substantial progress has been made in exploring auxin conjugation and metabolism, primarily aiming at indole-3-acetic acid (IAA). However, the metabolic regulation of another key auxin, phenylacetic acid (PAA), remains largely uncharacterized. Here, we provide a comprehensive exploration of PAA metabolism in land plants. Through LC-MS screening across multiple plant species and their organs, we identified four previously unreported endogenous PAA metabolites: phenylacetyl-leucine (PAA-Leu), phenylacetyl-phenylalanine (PAA-Phe), phenylacetyl-valine (PAA-Val), and phenylacetyl-glucose (PAA-glc). Enzyme assays, genetic evidence, crystal structures, and docking studies demonstrate that PAA and IAA share core metabolic machinery, revealing a complex regulatory network that maintains auxin homeostasis. Furthermore, our study of PAA conjugation with amino acids and glucose suggests limited compensatory mechanisms within known conjugation pathways, pointing to the existence of alternative metabolic routes in land plants. These insights advance our knowledge of auxin-specific metabolic networks and highlight the unique complexity within plant hormone regulation.

plant biology↗

bioRxiv2023

Cytokinin Dehydrogenase in Xylem Sap Reveals A Direct Link Between Cytokinin Metabolism and Long-Distance Transport

Metabolic degradation of plant hormones cytokinins (CKs) co-regulates their homeostasis and signalling. In this work, we employed a large-scale bioinformatical analysis to address a diversity of cytokinin oxidase/dehydrogenase (CKX) substrate specificities previously described in several case studies. We present a three-way correlation of the entire CKX amino acid sequences, a variable motif involved in substrate binding, and subcellular localizations predicted by a deep learning model. This correlation is conserved in monocotyledonous plants, suggesting that the CKX diversity in a single species allows a precise tuning of the CK homeostasis. Following these findings, we detected CKX activity in xylem sap for the first time, using the oat (Avena sativa) as a model plant. Further investigation of the substrate specificity and glycosylation of this xylem-located CKX suggested that it originates in roots. We have identified 27 putative CKXs in oats and attributed the xylem-located activity to the extracellular isoforms AsCKX1a,c,d. Finally, we show that the xylem-located CKX activity responds to the nitrate supply, highlighting its physiological relevance. Taken together, we show that CKX directly modulates root-to-shoot CK translocation through metabolic degradation of the transported CKs.

plant biology↗

bioRxiv2020

DIETARY MONOTERPENOIDS AS A NEW CLASS OF ALLOSTERIC HUMAN ARYL HYDROCARBON RECEPTOR ANTAGONISTS

Carvones, the constituents of essential oils of dill, caraway, and spearmint, were reported to antagonize the human aryl hydrocarbon receptor (AhR); however, the exact molecular mechanism remains elusive. We show that carvones are non-competitive allosteric antagonists of the AhR that inhibit the induction of AhR target genes in a ligand-selective and cell type-specific manner. Carvones do not displace radiolabeled ligand from binding at the AhR, but they bind allosterically within the bHLH/PAS-A region of the AhR. Carvones did not influence a translocation of ligand-activated AhR into the nucleus. Carvones inhibited the heterodimerization of the AhR with its canonical partner ARNT and subsequent binding of the AhR to the promotor of CYP1A1. Interaction of carvones with potential off-targets, including ARNT and protein kinases, was refuted. This is the first report of a small dietary monoterpenoids as a new class of AhR non-competitive allosteric antagonists with the potential preventive and therapeutic application.

pharmacology and toxicology↗