Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “PHOSPHATES”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 361 records · Page 20Linked to original sources
PubMed2002

Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway.

1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) catalyzes the first committed step of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis. In Arabidopsis, DXR is encoded by a single-copy gene. We have cloned a full-length cDNA corresponding to this gene. A comparative analysis of all plant DXR sequences known to date predicted an N-terminal transit peptide for plastids, with a conserved cleavage site, and a conserved proline-rich region at the N terminus of the mature protein, which is not present in the prokaryotic DXR homologs. We demonstrate that Arabidopsis DXR is targeted to plastids and localizes into chloroplasts of leaf cells. The presence of the proline-rich region in the mature Arabidopsis DXR was confirmed by detection with a specific antibody. A proof of the enzymatic function of this protein was obtained by complementation of an Escherichia coli mutant defective in DXR activity. The expression pattern of beta-glucuronidase, driven by the DXR promoter in Arabidopsis transgenic plants, together with the tissue distribution of DXR transcript and protein, revealed developmental and environmental regulation of the DXR gene. The expression pattern of the DXR gene parallels that of the Arabidopsis 1-deoxy-D-xylulose 5-phosphate synthase gene, but the former is slightly more restricted. These genes are expressed in most organs of the plant including roots, with higher levels in seedlings and inflorescences. The block of the 2-C-methyl-D-erythritol 4-phosphate pathway in Arabidopsis seedlings with fosmidomycin led to a rapid accumulation of DXR protein, whereas the 1-deoxy-D-xylulose 5-phosphate synthase protein level was not altered. Our results are consistent with the participation of the Arabidopsis DXR gene in the control of the 2-C-methyl-D-erythritol 4-phosphate pathway.

Aldose-Ketose Isomerases↗
PubMed2004

Direct demonstration of carbamoyl phosphate formation on the C-terminal domain of carbamoyl phosphate synthetase.

Carbamoyl phosphate synthetase synchronizes the utilization of two ATP molecules at duplicated ATP-grasp folds to catalyze carbamoyl phosphate formation. To define the dedicated functional role played by each of the two ATP sites, we have carried out pulse/labeling studies using the synthetases from Aquifex aeolicus and Methanococcus jannaschii, hyperthermophilic organisms that encode the two ATP-grasp folds on separate subunits. These studies allowed us to differentially label each active site with [gamma-(32)P]ATP and determine the fate of the labeled gamma-phosphate in the synthetase reaction. Our results provide the first direct demonstration that enzyme-catalyzed transfer of phosphate from ATP to carbamate occurs on the more C-terminal of the two ATP-grasp folds. These findings rule out one mechanism proposed for carbamoyl phosphate synthetase, where one ATP acts as a molecular switch, and provide additional support for a sequential reaction mechanism where the gamma-phosphate groups of both ATP molecules are transferred to reactants. CP synthesis by subunit C in our single turnover pulse/chase assays did not require subunit N, but subunit N was required for detectable CP synthesis in the traditional continuous assay. These findings suggest that cross-talk between domain N and C is required for product release from subunit C.

Adenosine Triphosphate↗
PubMed1975

Topology of binding sites for carbamyl phosphate in aspartate transcarbamylase from Escherichia coli. The use of pyridoxal phosphate as covalent probe.

Pyridoxal phosphate, a competitive inhibitor of aspartate transcarbamylase, binds to six sites in the catalytic and to twelve sites in the regulatory subunits of this hexameric protein. The properties of its association to the active sites of the enzyme are very similar to those observed with one of its substrates, carbamyl phosphate. It tightly binds to one half of the sites in the absence of succinate, an analogue of the second substrate. Since pyridoxal phosphate can be linked covalently to the protein by reduction, the distribution of the high affinity binding sites on catalytic trimers was studied after dissociation of modified holoenzyme. Electrophoresis of isolated subunits under non-denaturing conditions revealed four distinct bands, corresponding to trimers containing 0 to 3 pyridoxal phosphate derivatives. The distribution among the four species as a function of ligand concentration in the absence of succinate indicates that in the native oligomer, pyridoxal phosphate (and by extrapolation, carbamyl phosphate) binds to both catalytic trimers, rather than to three sites on a single subunit.

Aspartate Carbamoyltransferase↗
PubMed1974

Vitamin B6 metabolism in chronic alcohol abuse. Pyridoxal phosphate levels in plasma and the effects of acetaldehyde on pyridoxal phosphate synthesis and degradation in human erythrocytes.

The plasma pyridoxal-5'-phosphate (PLP) level of alcoholic subjects has been compared with that of non-alcoholic individuals in order to ascertain the incidence of abnormal vitamin B(6) metabolism in chronic alcohol abuse. 66 alcoholic subjects were selected on the basis that they did not exhibit abnormal liver function tests and hematologic findings. 35 of them had plasma PLP concentrations less than 5 ng/ml, the lowest value encountered in 94 control subjects, indicating a high incidence of deranged PLP metabolism in alcoholic patients even when hepatic and hematologic abnormalities are absent. The biochemical basis for the altered PLP metabolism in chronic alcohol abuse was examined. Low plasma PLP levels in alcoholics were not accompanied by decreased pyridoxal kinase and pyridoxine phosphate oxidase activities in erythrocytes. Further studies with erythrocytes demonstrated that the cellular content of PLP is determined not only by the activities of these PLP-synthesizing enzymes but also by the activity of a phosphate-sensitive, membrane-associated, neutral phosphatase, which hydrolyzes phosphorylated B(6) compounds.Acetaldehyde, but not ethanol, impaired the net formation of PLP from pyridoxal, pyridoxine, and pyridoxine phosphate by erythrocytes. However, when the B(6)-phosphate phosphatase activity was inhibited by 80 mM phosphate, this effect of acetaldehyde was abolished. By the use of broken cell preparations, it was possible to demonstrate directly that the action of acetaldehyde is mediated by the phosphatase, resulting in an acceleration of the degradation of the phosphorylated B(6) compounds in erythrocytes.

Acetaldehyde↗
PubMed1984

Mitochondrial phosphate transport. Large scale isolation and characterization of the phosphate transport protein from beef heart mitochondria.

The phosphate transport protein from beef heart mitochondria has been purified on a large scale by hydroxylapatite chromatography in the presence of sodium dodecyl sulfate and urea. As shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (silver stain), the pure phosphate transport protein preparation consists of two protein bands (alpha and beta, ratio 1:1) with similar mobilities (34 kDa) which display identical peptide maps if fragmented with either CNBr or HCl/dimethyl sulfoxide/HBr. The complete amino acid composition of phosphate transport protein is presented. Quantitative determination of N-terminal amino acids underlines the purity of the preparation and shows for alpha and beta the identical amino-terminals H2N-Ala-Val-Glu-Glu-Glx-Tyr-. Qualitative digestion shows that carboxypeptidase A is able to release at least three amino acids from the C termini of the alpha as well as the beta band of phosphate transport protein. The nature of these two protein bands is discussed. The sum of phosphate transport protein (alpha + beta) per total mitochondrial protein amounts to 2.3% or 1.4 nmol of phosphate transport protein (34 kDa) per nmol of cytochrome b.

Amino Acids↗
PubMed1982

pH gradient-dependent phosphate transport catalyzed by the purified mitochondrial phosphate transport protein.

The mitochondrial phosphate transport protein (Wohlrab, H. (1980) J. Biol. Chem. 255, 8170-8173), which co-purifies with the adenine nucleotide translocase, has been isolated with a modified procedure resulting in an increased yield and in a preparation that is stable to storage in liquid nitrogen. The phosphate transport protein was incorporated into liposomes (phosphatidylethanolamine/phosphatidylcholine/phosphatidic acid, 2.75:1:1), and phosphate/phosphate exchange rates were determined. With pHe (extraliposomal) = pHi (intraliposomal) = 7.2, we found a Km of 2.5 mM independent of [(Pi)i] and a Vmax of 12 mumol/min . mg of protein. Parallel phosphate transport experiments were carried out with liposomes containing phosphate transport protein isolated from mitochondria inhibited by N-ethylmaleimide. Phosphate transported unidirectionally [pHe = pHi = 6.8; = 1.0 mM, (Pi)i = 0 mM] reaches a maximum at 30 s and is 0.13 and 0.05 mumol/mg for active and inhibited protein, respectively. At pHe = 6.8 and pHi = 8.0, the respective amounts are 0.45 and 0.05. At pHe = 8.0, the uptake becomes the same with active and inhibited protein (pHi = 6.8, 0.15 and 0.14; pHi = 8.0, 0.25 and 0.20). At all the above pH values, about the same uptake is observed with liposomes prepared without protein as those prepared with inhibited protein. The initial rate of protein catalyzed unidirectional flux [(Pi)e = 1.0 mM, pHe = 6.8; (Pi)i = 0 mM, pHi = 8.0) is 2 mumol/min . mg of protein or 8 mumol/min . mg of phosphate transport protein estimated without the adenine nucleotide translocase.

Animals↗
PubMed1993

Effect of substrate and phosphate ions on the quaternary structure symmetry of glyceraldehyde-3-phosphate dehydrogenases of mung beans and rabbit muscle.

Effects of glyceraldehyde-3-phosphate (G-3-P) and phosphate ions on thermal inactivation of glyceraldehyde-3-phosphate dehydrogenases (GPDHs) of mung beans and rabbit muscle have been studied at different pH. In the absence of any ligand, the two enzymes show a striking similarity in the pH-dependence of the kinetics of thermal inactivation. At lower pH values both the enzymes biphasic kinetics with each phase accounting for about half of the starting activity (a C2 symmetry of the homotetrameric enzyme molecule). The kinetics change to a single exponential decay at higher pH values, a D2 symmetry [Malhotra & Srinivasan (1985) Arch. Biochem. Biphys. 236, 775-781; Malhotra & Tikoo (1991) Indian, J. Biochem. Biophys. 28, 16-21]. With each enzyme, phosphate ions are found to have no effect on the kinetic pattern at lower pH, but G-3-P brings about a change from biphasic to a single exponential decay. At higher pH values, G-3-P has no effect on the single exponential decay kinetic pattern, but phosphate ions change the same to a biphasic loss of activity with each phase accounting for about half of the starting activity. It has been concluded that with both the enzymes, G-3-P and phosphate ions have higher affinity and stabilise the D2- and C2-symmetry conformation, respectively. Binding isotherms of the two substrates for these enzymes have been described based on the ligand concentration-dependence of the changes in the rate constants and kinetic pattern of thermal inactivation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2004

A water setting tetracalcium phosphate-dicalcium phosphate dihydrate cement.

The development of a calcium phosphate cement, comprising tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD), that hardens in 14 min with water as the liquid or 6 min with a 0.25 mol/L sodium phosphate solution as the liquid, without using hydroxyapatite (HA) seeds as setting accelerator, is reported. It was postulated that reduction in porosity would increase cement strength. Thus, the effects of applied pressure during the initial stages of the cement setting reaction on cement strength and porosity were studied. The cement powder comprised an equimolar mixture of TTCP and DCPD (median particle sizes 17 and 1.7 microm, respectively). Compressive strengths (CS) of samples prepared with distilled water were 47.6 +/- 2.4 MPa, 50.7 +/- 4.2 MPa, and 52.9 +/- 4.7 MPa at applied pressures of 5 MPa, 15 MPa, and 25 MPa, respectively. When phosphate solution was used, the CS values obtained were 41.5 +/- 2.3 MPa, 37.9 +/- 1.7 MPa, and 38.1 +/- 2.3 MPa at the same pressure levels. Statistical analysis of the results showed that pressure produced an improvement in CS when water was used as liquid but not when the phosphate solution was used. Compared to previously reported TTCP-DCPD cements, the greater CS values and shorter setting times together with a simplified formulation should make the present TTCP-DCPD cement a useful material as a bone substitute for clinical applications.

Adhesives↗
PubMed2002

Different Mg to Fe ratios in the mixed metal MgFe hydroxy-carbonate compounds and the effect on phosphate binding compared with established phosphate binders.

Due to the side effects of the current oral phosphate binders, there is a need for effective alternatives. A number of mixed metal hydroxy-carbonate compounds (MMHCs) based on Mg and Fe have recently been established as effective phosphate binders. We have now carried out further studies on the MMHCs with different ratios of Mg(2+):Fe(3+) in different forms to assess for phosphate binding efficacy and ionic release in aqueous solution and food slurries. The compounds that provide the most promise are those with Mg(2+):Fe(3+) ratios of 2:1 and 4:1 in the unaged/dry form. Their phosphate binding efficacy was compared with a wide range of established phosphate binders, such as aluminum hydroxide [Al(OH)(3)], calcium carbonate (CaCO(3)), calcium acetate (CaAc(2)), magnesium hydroxide [Mg(OH)(2)], and lanthanum carbonate [La(2)(CO(3))(3)] in various food slurries. The results showed that the MgFe compounds were much more effective (on a weight for weight basis) than the established binders, and their properties were relatively pH independent. Calcium compounds (CaCO(3) and CaAc(2)) were ineffective under the experimental conditions. Mg(OH)(2) was effective at low pH but not at pHs greater than 5.0, and also released two- to threefold more magnesium than the MgFe compounds. Al(OH)(3) showed some degree of efficacy, but the binding capacity was, at best, less than 50% of the MMHCs. La(2)(CO(3))(3) required at least a 10-fold increase in weight to give comparable binding to the MMHCs. In conclusion, MgFe hydroxy-carbonate compounds are effective phosphate binders and may provide a better alternative to both existing and emerging binders for combating hyperphosphataemia.

Binding Sites↗
PubMed1987

Effect of alkaline-phosphatase inhibition by 1-p-bromotetramisole on the formation of trichloroacetic acid-[32P]-insoluble phosphate from inorganic [32P]-phosphate and [32P]-pyrophosphate in non-mineralizing and mineralizing hamster molar tooth-germs in vitro.

In culture, 1-p-bromotetramisole (pBTM), a specific inhibitor of alkaline phosphatase, significantly inhibited the formation of trichloroacetic acid (TCA)-insoluble [32P]-phosphate from inorganic [32P]-phosphate in the proliferating non-mineralizing second (M2) maxillary molar germs but had no effect in the actively mineralizing first (M1) germs. Addition of 10(-5) M inorganic pyrophosphate in the culture medium with a [32P]-phosphate label increased the inhibition of the formation of TCA-insoluble [32P]-phosphate in the M2. pBTM almost completely inhibited the formation of TCA-insoluble [32P]-phosphate from inorganic [32P]-pyrophosphate in the non-mineralizing M2. In the actively mineralizing M1, the compound significantly inhibited but did not abolish the formation of TCA-insoluble phosphate. These results confirm earlier biochemical findings that alkaline phosphatase possesses a pyrophosphatase activity probably related to the turnover of phosphorylated macromolecules necessary for cell differentiation and proliferation.

Alkaline Phosphatase↗
PubMed2004

Transport-deficient Pit2 phosphate transporters still modify cell surface oligomers structure in response to inorganic phosphate.

Pit2 is a member of the Pit family of inorganic phosphate transporters and serves as a gamma-retrovirus receptor in mammals. Pit2 contains two copies of the protein homology domain PD001131, which defines the Pit family. These domains are presumably in opposite topology with respect to the plasma membrane plane. We have mutated a serine residue conserved in almost all of the 192 known PD001131 sequences to alanine in each PD001131 domain of human Pit2. Expression in CHO cells showed that phosphate uptake was affected severely in mutants, whereas susceptibility to virus infection was conserved. We reported previously that the inorganic phosphate concentration affects both phosphate transport mediated by Pit2 and the conformation of cell-surface Pit2 oligomers. Cross-linking experiments in transport-incompetent Pit2 mutants indicated that structural changes induced by phosphate starvation or supply occur independently of the whole transport cycle. These results suggest that the structural reorganisation of cell-surface Pit2 occurred as a consequence of ion binding, a model consistent with the possible involvement of cell-surface Pit2 oligomers in inorganic phosphate sensing.

Amino Acid Sequence↗
PubMed2004

Effect of nitrite on phosphate uptake by phosphate accumulating organisms.

In biological nitrogen removal processes, nitrite can be formed and accumulated through both nitrification and denitrification. Despite the fact that, in practice, biological phosphate removal (BPR) is often combined with biological nitrogen removal, there are only a few publications reporting the effect of nitrite on BPR. In this study, phosphate-accumulating organisms (PAO) were cultivated in an anaerobic-anoxic-aerobic sequencing batch reactor (SBR). The effect of nitrite on the enrichment of the sludge with PAO, the phosphate uptake rates and the sludge respiration was investigated. The results indicate that (1) presence of nitrite inhibits both aerobic and anoxic (denitrifying) phosphate uptake, (2) aerobic phosphate uptake was more affected than anoxic phosphate uptake, (3) presence of nitrite could be one of the factors enhancing the presence of glycogen accumulating organisms (GAO)--competitors to PAO for substrate in the anaerobic phase, and (4) it is required to monitor and control nitrite accumulation in a full-scale wastewater treatment plants.

Bacteria, Anaerobic↗
PubMed2000

Serum phosphate in stone formers: what does a low serum phosphate level mean?

PURPOSE: Metabolic evaluation is indicated in stone formers to identify and subsequently modify risk factors for nephrolithiasis. Some believe that serum phosphate may be an important indicator of nephrolithiasis risk. We determined whether serum phosphate is an independent risk factor for recurrent calcium stone formation and whether it may be used as an early marker for occult disease, such as hyperparathyroidism. MATERIALS AND METHODS: The charts of all patients with regular long-term stone clinic followup were reviewed. Initial serum phosphate levels were compared with stone recurrence, complications due to stones, initial visit urinary laboratory values and the medical diagnoses made during followup. Unpaired 2-sample t tests, and correlation and logistic regression analyses were performed with statistical significance at p <0.05. RESULTS: Data were available on 51 men and 25 women 22 to 74 years old (mean age 52). Of the patients 50 were recurrent and 26 were single stone formers. Serum phosphate levels were lower in women than in men. No consistent associations existed between initial or future serum phosphate levels and stone recurrence, complications from stones, urinary values or other medical diagnoses made during followup. CONCLUSIONS: Serum phosphate does not appear to be an independent risk factor for urinary tract stone recurrence or complications, or a reliable early predictor of occult disease.

Adult↗
PubMed1978

Phosphate content of Escherichia coli alkaline phosphatase isozymes. The effect of phosphate and zinc on the separation of isozymes.

Alkaline phosphatase from Escherichia coli was isolated as two major isoenzyme forms that were separated by DEAE-cellulose chromatography. Each form contained 2 equiv of endogenous phosphate. The endogenous phosphate, although difficult to remove, readily exchanges with phosphate. The forms also were separable by polyacrylamide gel electrophoresis. Apoenzyme prepared from native enzyme by the removal of zinc (and phosphate) also contains electrophoretically distinct enzyme forms which are indistinguishable from the native forms on gel electrophoresis. The isozymes were also found to have similar affinities for inorganic phosphate and susceptibilities to inactivation by EDTA. These results are not consistent with the notion that the formation or separation of isoenzyme forms is dependent upon different amounts of bound phosphate. They are consistent with the suggestion that a difference in amino acid composition is the basis for the occurrence and separation of these isoenzymes.

Alkaline Phosphatase↗
PubMed2004

Enhanced accumulation of phosphate by Lolium multiflorum cultivars grown in phosphate-enriched medium.

Agricultural and animal husbandry practices combined with soil composition have caused phosphate overloading of farmlands in different parts of the U.S. and Europe. Movement of soluble phosphates (Pi) from phosphorus enriched soils results in degradation of natural aquatic systems, triggering serious environmental problems. Remediation of such sites using plants that tolerate and accumulate high concentrations of Pi in their aerial parts may be an attractive remediation technology. In the present study, Pi transport and accumulation potential of Marshall and Gulf ryegrass (Lolium multiflorum cultivars) were determined using a solution culture of seedlings. Ryegrass seedlings accumulated phosphorus (P) in excess of 2% of dry weight in their aerial parts when supplied with 5 g/L KH2PO4 in medium. Phosphorus accumulation was positively correlated with the concentration of phosphate (0-5 g/L KH2PO4) in medium. Plants grew well on medium containing 5 g/L KH2PO4, but concentrations above 5 g/L caused symptoms of toxicity. Scanning electron microscopy and energy-dispersive X-ray spectroscopy confirmed high P accumulation in different cell types of grass roots and shoots. Phosphate starvation and replenishment experiments point to the unique ability of these grasses to concentrate phosphate in the above-ground parts. It is hypothesized that the unique ability of these ryegrass cultivars may be due to the presence of efficient phosphate transport and sequestration mechanisms.

Agriculture↗
PubMed2003

1H NMR titrations of hydroxy protons in aqueous solution as a method of investigation of intramolecular hydrogen-bonding in phosphorylated compounds: examples of myo-inositol 2-phosphate and myo-inositol 1,2,6-tris(phosphates).

1H NMR hydroxy proton titration experiments for myo-inositol 2-phosphate and myo-inositol 1,2,6-tris(phosphates) in aqueous solution are presented to demonstrate that by following OH signals versus pH, evidence can be brought for HB interaction between the hydroxyl and phosphate groups. The chemical shifts of the OH protons vicinal to phosphate groups appear deshielded by ca. 2.5 ppm with regard to those two centers removed from the phosphates. Remarkably, the deshielded protons are only present when their neighboring phosphate groups are fully deprotonated but persist until high pHs. From these results, C-OH...2-O3P-O type I, C-HO...-HO3P-O type II and C-OH...-HO3P-O type III hydrogen bonds are evidenced and discussed.

Hydrogen Bonding↗
PubMedDate not supplied

Dihydroxyacetone, pyruvate, and phosphate effects on 2,3 DPG and ATP in citrate-phosphate-dextrose-adenine blood preservation.

Blood was stored with various combinations of 60 mM dihydroxyacetone (DHA), 80 mM pyruvate and 10 mM phosphate in citrate-phosphate-dextrose-adenine-one (CPDA-1) preservative for 42 days. During that time, DHA was found to have a beneficial effect on 2,3 diphosphoglycerate (2,3 DPG), although a deleterious effect on adenosine triphosphate (ATP). Preservatives containing pyruvate gave similar results. However, DHA and phosphate together in CPDA-1 improved both 2,3 DPG and ATP. The adverse effect on 2,3 DPG levels that phosphate normally shows was reversed by the addition of DHA. These results indicate that the addition of phosphate alone, rather than pyruvate, or pyruvate and phosphate, to DHA in CPDA-1 would be preferable to the presently licensed CPDA-1 preservative.

2,3-Diphosphoglycerate↗
PubMed1999

Crystalline phase differentiation in urinary calcium phosphate and magnesium phosphate calculi.

UNLABELLED: Phosphates are encountered as the main components in about 15% of urinary calculi. Except for struvite, no specific correlations have been found between the crystalline phase of the phosphates and the cause of nephrolithiasis. OBJECTIVE: The relationship between aetiological factors and crystalline phases or carbonate rate in calcium phosphate stones were assessed. MATERIAL AND METHODS: From a series of 1148 phosphate calculi, we investigated the relationship between composition and aetiological factors. RESULTS: Carbapatite was the most frequent crystalline phase (74.0%). It was associated with many possible causes, including hypercalciuria, hypocitraturia, primary hyperparathyroidism, tubular acidosis, medullary sponge kidney and chronic urinary tract infection. The carbonate rate of carbapatite may be of clinical interest because carbonate rates above 15% are frequently related to urinary tract infection with urea-splitting bacteria. Conversely, the carbonate rate was commonly less than 10% in cases of carbapatite induced by metabolic disorders. Among other phosphates, brushite was found in hypercalciuric states and primary hyperparathyroidism and whitlockite in cases of urinary tract infection by non-urease-producing bacteria. CONCLUSION: Identification of crystalline phases and measurement of carbonate rate in calcium phosphate calculi is of clinical interest for identifying stone aetiology.

Acidosis↗