The phytase RipBL1 enables the assignment of a specific inositol phosphate isomer as a structural component of human kidney stones

Liu, Guizhen; Riemer, Esther; Schneider, Robin; Cabuzu, Daniela; Bonny, Olivier; Wagner, Carsten A.; Qiu, Danye; Saiardi, Adolfo; Strauß, Annett; Lahaye, Thomas; Schaaf, Gabriel; Knoll, Thomas; Jessen, Jan Peter; Jessen, Henning J.

doi:10.1039/d2cb00235c

Cited by 14 publications

(10 citation statements)

References 58 publications

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“…To this end, inositol phosphates from the thallus of Marchantia polymorpha (liverworts; Bryophyta), sporophylls of Selaginella sp. (Lycopods; Pteridophyta) and seedlings of Arabidopsis thaliana (Eudicot, Angiosperms) were extracted and analyzed using the recently developed capillary electrophoresis coupled with mass spectrometry method (CE-MS)(Liu et al, 2023; Qiu et al, 2023; Qiu et al, 2020). Our analyses demonstrated the presence of 4/6-InsP 7 in all plant extracts used in the study (Fig.…”

Section: Resultsmentioning

confidence: 99%

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP₇production in land plants

Yadav,

Liu,

Rana

et al. 2023

Preprint

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Inositol pyrophosphates (PP-InsPs) are soluble cellular messengers that integrate environmental cues to induce adaptive responses in eukaryotes. In plants, the biological functions of various PP-InsP species are poorly understood, largely due to the absence of canonical enzymes present in other eukaryotes. The recent identification of a new PP-InsP isomer with yet unknown enantiomeric identity, 4/6-InsP7in the eudicotArabidopsis thaliana, further highlights the intricate PP-InsP signalling network employed by plants. The abundance of 4/6-InsP7in land plants, the enzyme(s) responsible for its synthesis, and the physiological functions of this species are all currently unknown. In this study, we show that 4/6-InsP7is the major PP-InsP species present across land plants. Our findings demonstrate that theArabidopsisinositol polyphosphate multikinase (IPMK) homolog, AtIPK2α generates 4/6-InsP7in vitro. Furthermore, the cellular level of 4/6-InsP7is controlled by the twoArabidopsisIPMK isoforms, AtIPK2α and AtIPK2β. Notably, the activity of these IPMK proteins is critical for heat stress acclimation inArabidopsis. During heat stress, the expression of genes encoding various heat shock proteins controlled by the heat shock factors (HSFs) is affected in the AtIPK2-deficient plants. Furthermore, we show that the transcription activity of HSF is regulated by the AtIPK2 proteins. Our parallel investigations using the liverwortMarchantia polymorphasuggest that the InsP6kinase activity of IPMK and the role of IPMK in regulating the heat stress response are evolutionarily conserved. Collectively, our study indicates that IPMK has played a critical role in transducing environmental cues for biological processes during land plant evolution.

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Section: Resultsmentioning

confidence: 99%

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP₇production in land plants

Yadav,

Liu,

Rana

et al. 2023

Preprint

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“…The salient advantages of CE are high separation efficiencies in combination with nanoliter sample consumption and low operating costs. Importantly, the TiO 2 extraction protocol will now enable to correlate ppGpp and InsP signaling in plants by parallel determinations, which will potentially uncover cross-talk between these pathways. ,,− Each analysis presented herein is based on a 20 nL injection volume of the analyte solution, while LC-MS approaches require 50 to 500 times more sample volume (1–10 μL). , Determination of the limit of detection (LOD) and limit of quantification (LOQ) were achieved via spiking of extracted plant material (150 mg FW) with heavy internal standards and calculation of a signal-to-noise ratio above 3 (LOD) and 10 (LOQ). The LOD for ppGpp expressed as a concentration was 10 nM, and the LOQ was 30 nM in this particular matrix (see Supporting Figures S5 and S6).…”

Section: Resultsmentioning

confidence: 99%

Bacterial Pathogen Infection Triggers Magic Spot Nucleotide Signaling in Arabidopsis thaliana Chloroplasts through Specific RelA/SpoT Homologues

et al. 2023

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Magic spot nucleotides (p)ppGpp are important signaling molecules in bacteria and plants. In the latter, RelA-SpoT homologue (RSH) enzymes are responsible for (p)ppGpp turnover. Profiling of (p)ppGpp is more difficult in plants than in bacteria due to lower concentrations and more severe matrix effects. Here, we report that capillary electrophoresis mass spectrometry (CE-MS) can be deployed to study (p)ppGpp abundance and identity in Arabidopsis thaliana. This goal is achieved by combining a titanium dioxide extraction protocol and pre-spiking with chemically synthesized stable isotope-labeled internal reference compounds. The high sensitivity and separation efficiency of CE-MS enables monitoring of changes in (p)ppGpp levels in A. thaliana upon infection with the pathogen Pseudomonas syringae pv. tomato (PstDC3000). We observed a significant increase of ppGpp post infection that is also stimulated by the flagellin peptide flg22 only. This increase depends on functional flg22 receptor FLS2 and its interacting kinase BAK1 indicating that pathogen-associated molecular pattern (PAMP) receptor-mediated signaling controls ppGpp levels. Transcript analyses showed an upregulation of RSH2 upon flg22 treatment and both RSH2 and RSH3 after PstDC3000 infection. Arabidopsis mutants deficient in RSH2 and RSH3 activity display no ppGpp accumulation upon infection and flg22 treatment, supporting the involvement of these synthases in PAMP-triggered innate immune responses to pathogens within the chloroplast.

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“…The accurate analysis and identification of regioisomers and enantiomers is a significant challenge but crucial for understanding the complex network of InsPs and PP-InsPs in biological systems [9,23,37,38]. Enantiomer assignment has traditionally been accomplished using crystal structures, enzymatic assays in combination with chemically synthesized standard compounds, and isotopically labeled myo-inositol (1) in combination with 13 C-, 31 P-, and two-dimensional-NMR measurements [14,15,[18][19][20][21][22][23].…”

Section: Discussionmentioning

confidence: 99%

Assigning the Absolute Configuration of Inositol Poly- and Pyrophosphates by NMR Using a Single Chiral Solvating Agent

Ritter,

Jork,

Unmüßig

et al. 2023

Biomolecules

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Inositol phosphates constitute a family of highly charged messenger molecules that play diverse roles in cellular processes. The various phosphorylation patterns they exhibit give rise to a vast array of different compounds. To fully comprehend the biological interconnections, the precise molecular identification of each compound is crucial. Since the myo-inositol scaffold possesses an internal mirror plane, enantiomeric pairs can be formed. Most commonly employed methods for analyzing InsPs have been geared towards resolving regioisomers, but they have not been capable of resolving enantiomers. In this study, we present a general approach for enantiomer assignment using NMR measurements. To achieve this goal, we used 31P-NMR in the presence of L-arginine amide as a chiral solvating agent, which enables the differentiation of enantiomers. Using chemically synthesized standard compounds allows for an unambiguous assignment of the enantiomers. This method was applied to highly phosphorylated inositol pyrophosphates, as well as to lowly phosphorylated inositol phosphates and bisphosphonate analogs. Our method will facilitate the assignment of biologically relevant isomers when isolating naturally occurring compounds from biological specimens.

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The phytase RipBL1 enables the assignment of a specific inositol phosphate isomer as a structural component of human kidney stones

Abstract: Inositol phosphates (InsPs) are ubiquitous in all eukaryotes. However, since there are 63 possible different phosphate ester isomers, the analysis of InsPs is challenging. In particular, InsP1, InsP2 and InsP3...

Cited by 14 publications

References 58 publications

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP₇production in land plants

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP₇production in land plants

Bacterial Pathogen Infection Triggers Magic Spot Nucleotide Signaling in Arabidopsis thaliana Chloroplasts through Specific RelA/SpoT Homologues

Assigning the Absolute Configuration of Inositol Poly- and Pyrophosphates by NMR Using a Single Chiral Solvating Agent

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The phytase RipBL1 enables the assignment of a specific inositol phosphate isomer as a structural component of human kidney stones

Abstract: Inositol phosphates (InsPs) are ubiquitous in all eukaryotes. However, since there are 63 possible different phosphate ester isomers, the analysis of InsPs is challenging. In particular, InsP1, InsP2 and InsP3...

Cited by 14 publications

References 58 publications

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP7production in land plants

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP7production in land plants

Bacterial Pathogen Infection Triggers Magic Spot Nucleotide Signaling in Arabidopsis thaliana Chloroplasts through Specific RelA/SpoT Homologues

Assigning the Absolute Configuration of Inositol Poly- and Pyrophosphates by NMR Using a Single Chiral Solvating Agent

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Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP₇production in land plants

Conservation of heat stress acclimation by the inositol polyphosphate multikinase, IPMK responsible for 4/6-InsP₇production in land plants