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Inositol phosphates are signaling molecules crucial for cellular processes. To achieve specificity, their diverse phosphorylation patterns necessitate precise molecular identification. Phosphorylation of the myo-inositol scaffold can form enantiomeric pairs, but most analysis methods only resolve regioisomers. This chapter presents an approach for enantiomer assignment using P-NMR with L-arginine amide as a chiral solvating agent. Chemically synthesized standards enabled clear enantiomer differentiation. This method applies to both inositol phosphates as well as inositol pyrophosphates and their bisphosphonate analogs, aiding in the identification of biologically relevant isomers in natural samples.
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http://dx.doi.org/10.1007/978-1-0716-4799-8_9 | DOI Listing |
Nat Commun
September 2025
Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Oxford, UK.
Inositol hexakisphosphate (IP6) promotes HIV-1 assembly by stabilizing the immature Gag lattice and becomes enriched within virions, where it is required for mature capsid assembly. Previously, we identified Gag mutants that package little IP6 yet assemble particles, though they are non-infectious due to defective capsid formation. Here, we report a compensatory mutation, G225R, in the C-terminus of capsid protein (CA) that restores capsid assembly and infectivity in these IP6-deficient mutants.
View Article and Find Full Text PDFBiomolecules
August 2025
Renal Lithiasis and Pathological Calcification Group, Research Institute of Health Sciences (IUNICS), University of the Balearic Islands, 07122 Palma, Spain.
Thermodynamic factors (supersaturation of substances that form crystals) and kinetic factors (heterogeneous nucleants and crystallization inhibitors) affect the formation of crystals and stones in the urinary tract. We studied the effect of five different polyhydroxycarboxylic acids and phytate on the formation of calcium oxalate crystals in artificial urine. All tested molecules are known to inhibit the crystallization of this calcium salt, and to also form complexes with calcium ions.
View Article and Find Full Text PDFMethods Mol Biol
August 2025
Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD, USA.
Inositol pyrophosphates, such as 5-diphosphoinositol pentakisphosphate (IP7), play crucial roles in various biological processes. Pharmacologic inhibition of IP6K has potential therapeutic benefits for treating type II diabetes, venous thrombosis, chronic kidney disease, and psychiatric disorders. This chapter describes the identification of IP6K inhibitors through kinase-activity-based high-throughput assays and dose-response assay.
View Article and Find Full Text PDFMethods Mol Biol
August 2025
Laboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics, Hyderabad, India.
Knockout mice have served as excellent model systems to investigate the functions of specific mammalian proteins at the organismal level. Several studies on tissue-specific or whole-body depletion of individual IP6 kinase (IP6K) paralogs have shed light on myriad roles for their product, the inositol pyrophosphate 5-InsP, in different physiological processes and pathological states. The loss of Ip6k1, Ip6k2, or Ip6k3 leads predominantly to nonoverlapping phenotypes in mice, reflecting their differential tissue, cell type, and subcellular distribution.
View Article and Find Full Text PDFMethods Mol Biol
August 2025
Department of Plant Nutrition, Institute of Crop Science and Resource Conservation (INRES), University of Bonn, Bonn, Germany.
In recent years, there has been substantial progress in the development of methods to analyze inositol phosphates (InsPs) and inositol pyrophosphates (PP-InsPs). However, many of these techniques are labor- and cost-intensive and can usually only be carried out by laboratories specialized in InsPs/PP-InsPs analysis. In this chapter, we present a simple method that exploits the fact that phosphorylation and/or dephosphorylation of certain InsP/PP-InsP species induces the activation of promoters driving the expression of genes involved in phosphate starvation response (PSR).
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