The ability of bone-marrow-derived mesenchymal stem/stromal cells (BM-MSCs) to differentiate into osteoblasts makes them the ideal candidate for cell-based therapies targeting bone-diseases. ...Polyphosphate (polyP) is increasingly being studied as a potential inorganic source of phosphate for extracellular matrix mineralisation. The aim of this study is to investigate whether polyP can effectively be used as a phosphate source during the in vitro osteogenic differentiation of human BM-MSCs. Human BM-MSCs are cultivated under osteogenic conditions for 28 days with phosphate provided in the form of organic β-glycerolphosphate (BGP) or calcium-polyP nanoparticles (polyP-NP). Mineralisation is demonstrated using Alizarin red staining, cellular ATP content, and free phosphate levels are measured in both the cells and the medium. The effects of BGP or polyP-NP on alkaline phosphatase (ALP) activity and gene expression of a range of osteogenic-related markers are also assessed. PolyP-NP supplementation displays comparable effects to the classical BGP-containing osteogenic media in terms of mineralisation, ALP activity and expression of osteogenesis-associated genes. This study shows that polyP-NP act as an effective source of phosphate during mineralisation of BM-MSC. These results open new possibilities with BM-MSC-based approaches for bone repair to be achieved through doping of conventional biomaterials with polyP-NP.
Persistence refers to the phenomenon in which isogenic populations of antibiotic-sensitive bacteria produce rare cells that transiently become multidrug tolerant. Whether slow growth in a rare subset ...of cells underlies the persistence phenotype has not be examined in wild-type bacteria. Here, we show that an exponentially growing population of wild-type Escherichia coli cells produces rare cells that stochastically switch into slow growth, that the slow-growing cells are multidrug tolerant, and that they are able to resuscitate. The persistence phenotype depends hierarchically on the signaling nucleotide (p)ppGpp, Lon protease, inorganic polyphosphate, and toxin-antitoxins. We show that the level of (p)ppGpp varies stochastically in a population of exponentially growing cells and that the high (p)ppGpp level in rare cells induces slow growth and persistence. (p)ppGpp triggers slow growth by activating toxin-antitoxin loci through a regulatory cascade depending on inorganic polyphosphate and Lon protease.
The TRPA1 ion channel (also known as the wasabi receptor) is a detector of noxious chemical agents encountered in our environment or produced endogenously during tissue injury or drug metabolism. ...These include a broad class of electrophiles that activate the channel through covalent protein modification. TRPA1 antagonists hold potential for treating neurogenic inflammatory conditions provoked or exacerbated by irritant exposure. Despite compelling reasons to understand TRPA1 function, structural mechanisms underlying channel regulation remain obscure. Here we use single-particle electron cryo- microscopy to determine the structure of full-length human TRPA1 to ∼4 Å resolution in the presence of pharmacophores, including a potent antagonist. Several unexpected features are revealed, including an extensive coiled-coil assembly domain stabilized by polyphosphate co-factors and a highly integrated nexus that converges on an unpredicted transient receptor potential (TRP)-like allosteric domain. These findings provide new insights into the mechanisms of TRPA1 regulation, and establish a blueprint for structure-based design of analgesic and anti-inflammatory agents.
Glutamine synthetase (GS) is vital in maintaining ammonia and glutamate (Glu) homeostasis in living organisms. However, the natural enzyme relies on adenosine triphosphate (ATP) to activate Glu, ...resulting in impaired GS function during ATP-deficient neurotoxic events. To date, no reports demonstrate using artificial nanostructures to mimic GS function. In this study, we synthesize aggregation-induced emission active polyP-Mn nanosheets (STPE-PMNSs) based on end-labeled polyphosphate (polyP), exhibiting remarkable GS-like activity independent of ATP presence. Further investigation reveals polyP in STPE-PMNSs serves as phosphate source to activate Glu at low ATP levels. This self-feeding mechanism offers a significant advantage in regulating Glu homeostasis at reduced ATP levels in nerve cells during excitotoxic conditions. STPE-PMNSs can effectively promote the conversion of Glu to glutamine (Gln) in excitatory neurotoxic human neuroblastoma cells (SH-SY5Y) and alleviate Glu-induced neurotoxicity. Additionally, the fluorescence signal of nanosheets enables precise monitoring of the subcellular distribution of STPE-PMNSs. More importantly, the intracellular fluorescence signal is enhanced in a conversion-responsive manner, allowing real-time tracking of reaction progression. This study presents a self-sustaining strategy to address GS functional impairment caused by ATP deficiency in nerve cells during neurotoxic events. Furthermore, it offers a fresh perspective on the potential biological applications of polyP-based nanostructures.
Summary
Short chain polyphosphates (polyP) are pro-coagulant and pro-inflammatory platelet released inorganic polymers. The platelet chemokine platelet factor 4 (PF4) binds to lipid A on bacteria, ...inducing an antibody mediated host defense mechanism, which can be misdirected against PF4/heparin complexes leading to the adverse drug reaction heparin-induced thrombocytopenia (HIT). Here, we demonstrate that PF4 complex formation with soluble short chain polyP contributes to host defense mechanisms. Circular dichroism spectroscopy and isothermal titration calorimetry revealed that PF4 changed its structure upon binding to polyP in a similar way as seen in PF4/heparin complexes. Consequently, PF4/polyP complexes exposed neoepitopes to which human anti-PF4/heparin antibodies bound. PolyP enhanced binding of PF4 to
Escherichia coli
, hereby facilitating bacterial opsonisation and, in the presence of human anti-PF4/polyanion antibodies, phagocytosis. Our study indicates a role of polyP in enhancing PF4-mediated defense mechanisms of innate immunity.
Inorganic polyphosphate (polyP) is produced by both bacteria and their eukaryotic hosts, and it appears to play multiple important roles in the interactions between those organisms. However, the ...detailed mechanisms of how polyP synthesis is regulated in bacteria, and how it influences both bacterial and host biology, remain largely unexplored. In this review, we examine recent developments in the understanding of how bacteria regulate the synthesis of polyP, what roles polyP plays in controlling virulence in pathogenic bacteria, and the effects of polyP on the mammalian immune system, as well as progress on developing drugs that may be able to target bacterial polyP synthesis as novel means of treating infectious disease.
Inorganic polyphosphate (polyP) plays multiple roles in bacterial regulation, including controlling proteolysis by the Lon protease and regulating virulence factors, both positively and negatively.PolyP plays a major role in host repair by facilitating thrombin-driven coagulation and in defense mechanisms by recruiting neutrophils and driving M1 macrophage differentiation.Bacteria use polyP to manipulate host immune responses, impairing phagocytic cell functions, modulating inflammatory responses, and impairing antigen presentation.Dictyostelium discoideum is emerging as a powerful model system for investigating polyP biology.Multiple drugs influencing polyP levels in different organisms have recently been discovered.
Phosphate is mined from phosphate rock, which is a limited resource on a human time scale. For a sustainable phosphate supply, strategies for efficient use and recycling of phosphate must be ...developed. A German chemical company produces annually wash water containing phosphate and other inorganic substances (e.g., sodium, potassium, sulfate, and chloride) at a ton scale. Chemical precipitation is mostly used for phosphate removal. In this study, a biotechnological process utilizing Saccharomyces cerevisiae to upcycle phosphate‐containing wastewater into pure sodium polyphosphate in powder form was developed. The process comprises fermentation and downstream processing (polyphosphate yields: 25% and 36%, respectively). The polyphosphate quality was independent of the wash water composition. Polyphosphate with a purity of 23% molar ratio Na to Na, K, and Mg of > 90%, and with an average chain length of 12.5 phosphate subunits was produced. The upcycled polyphosphate can be reused compared to phosphate fertilizer in many different applications. Overall, the here developed process can contribute to truly slowing down phosphate mining and finally enable a sustainable utilization of phosphate. Thereby, the benefit of the process is the cascade use of phosphate, reducing the need for phosphate rock before the phosphate ends up in the soil and ultimately in the sea.
The biotechnological production of pure sodium polyphosphate by Saccharomyces cerevisiae utilizing phosphate‐containing industrial wash water is described. The authors compared the polyphosphate production with different phosphate‐rich wash waters and were able to show that the polyphosphate quality was independent of the wash water composition.
Tetrasphaera and Candidatus Accumulibacter are two abundant polyphosphate accumulating organisms in full-scale enhanced biological phosphorus removal (EBPR) systems. However, little is known about ...the metabolic behaviour and ecological niche that each organism exhibits in mixed communities. In this study, an enriched culture of Tetrasphaera and Ca. Accumulibacter was obtained using casein hydrolysate as sole carbon source. This culture was able to achieve a high phosphorus removal efficiency (>99%), storing polyphosphate while consuming amino acids anaerobically. Microautoradiography and fluorescence in situ hybridisation confirmed that more than 90% Tetrasphaera cells were responsible for amino acid consumption while Ca. Accumulibacter likely survived on fermentation products. Tetrasphaera performed the majority of the P removal (approximately 80%) in this culture, and batch tests showed that the metabolism of some carbon sources could actually lead to anaerobic orthophosphate (Pi) uptake (9.0 ± 2.1 mg-P/L) through energy generated by fermentation of glucose and amino acids. This anaerobic Pi uptake may lead to lower net Pi release to C uptake ratios and reduce the Pi needed to be removed aerobically in WWTPs. Intracellular metabolites such as amino acids, sugars, volatile fatty acids and small amines were observed as potential storage products, which may serve as energy sources in the aerobic phase. Evidence of the urea cycle was found, which could be involved in reducing the intracellular nitrogen content. This study improves our understanding of how phosphorus is removed in EBPR systems and can enable novel process optimisation strategies.
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•A P-removing, enriched Tetrasphaera and Ca. Accumulibacter culture was obtained.•Tetrasphaera were mainly responsible for amino acid biodegradation and P removal.•Tetrasphaera are capable of anaerobic P uptake through energy from fermentation.•Numerous intracellular metabolites were identified - could serve as storage products.•Improved metabolic understanding could help optimise EBPR systems.
The pandemic caused by the novel coronavirus SARS-CoV-2 is advancing rapidly. In particular, the number of severe courses of the disease is still dramatically high. An efficient drug therapy that ...helps to improve significantly the fatal combination of damages in the airway epithelia, in the extensive pulmonary microvascularization and finally multiorgan failure, is missing. The physiological, inorganic polymer, polyphosphate (polyP) is a molecule which could prevent the initial phase of the virus life cycle, the attachment of the virus to the target cells, and improve the epithelial integrity as well as the mucus barrier.
Surprisingly, polyP matches perfectly with the cationic groove on the RBD. Subsequent binding studies disclosed that polyP, with a physiological chain length of 40 phosphate residues, abolishes the binding propensity of the RBD to the ACE2 receptor. In addition to this first mode of action of polyP, this polymer causes in epithelial cells an increased gene expression of the major mucins in the airways, of
and
, as well as a subsequent glycoprotein production. MUC5AC forms a gel-like mucus layer trapping inhaled particles which are then transported out of the airways, while MUC1 constitutes the periciliary liquid layer and supports ciliary beating. As a third mode of action, polyP undergoes enzymatic hydrolysis of the anhydride bonds in the airway system by alkaline phosphatase, releasing metabolic energy.
This review summarizes the state of the art of the biotherapeutic potential of the polymer polyP and the findings from basic research and outlines future biomedical applications.
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•It should cautiously pursue high PRA when aiming at improving PRE.•PRE plateaued at 75.68% with influent C/P ratio of 40 mgCOD/mgP.•Ca. Competibacter and Defluviicoccus were ...unexpectedly highly enriched.•Ca. Accumulibacter and Tetrasphaera were found absented from EBPR.
Competition between polyphosphate- and glycogen-accumulating organisms (PAOs and GAOs) is problematic in the enhanced biological phosphorus removal (EBPR) process. Aiming at a high phosphorus removal efficiency (PRE), the phosphorus release amount (PRA) is considered an essential evaluating indicator. However, the correlations between PRE and PRA and the abundance of PAOs are not clear. In this study, the EBPR was established and optimized via adjusting influent carbon to phosphorus ratio (C/P). After 110-day operation, 17.67 mg/L of PRA and 75.86% of PRE simultaneously achieved with influent C/P of 40 mgCOD/mgP. As for PAOs, Candidatus_Accumulibacter and Tetrasphaera were absent, while Hypomicrobium (3.69%), Pseudofulvimonas (1.02%), and unclassified_f_Rhodobacteraceae (2.41%) were found at a low level. On the contrary, Candidatus_Competibacter and Defluviicoccus were unexpectedly enriched with high abundance (24.94% and 16.04%, respectively). These results also suggested that it was difficult to distinguish whether PAOs were enriched merely based on the variations of PRA and PRE.
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