Water-soluble Phosphate Research Articles

Water-soluble phosphates of fish red cells

The nature and significance of water-soluble (small molecule) phosphates in vertebrate red blood cells are reviewed and new data presented for several fish including: three salmon (Oncorhynchus gorbuscha, Oncorhynchus kisutch and Oncorhynchus nerka), a mackerel (Scomber japonicus), a dogfish (Squalus acanthias), the ratfish (Hydrolagus colliei), and two air-breathing fish (Synbranchus marmoratus and Pterygoblichthys) studied on the research vessel Alpha Helix on the Amazon River. Of special interest was the wide variation in the ratio of guanosine triphosphate (GTP) to adenosine triphosphate (ATP); this ratio was much higher in some species of fish than had been reported for red cells of other vertebrates or, for that matter, for any other kind of tissue. The GTP may function in some fish as a regulator of haemoglobin–oxygen affinity.

Red blood cell lysis induced by the venom of the brown recluse spider: The role of sphingomyelinase D

Red blood cell lysis induced by the venom of Loxosceles reclusa, the brown recluse spider, may be related to the hemolytic anemia observed in several cases of spider envenomation. These investigations demonstrate that the venom of the brown recluse spider contains a calcium-dependent, heat-labile hemolysin of molecular weight approximately 19,000. The pH optimum for the hemolytic reaction was 7.1, and the optimum calcium concentration for venom-induced lysis was observed within the range of 6 to 10 m m. Sheep red blood cells were more susceptible to the spider hemolysin than human red blood cells, although both types exhibited appreciable lysis. Digestion of sheep red blood cell membranes with partially purified venom lysin resulted in degradation of the sphingomyelin component. However, reaction of the membranes with the venom lysin produced no release of water-soluble phosphate, and no free fatty acids were generated. These results indicate that the sphingomyelin-degrading activity of the venom is not a phospholipase C- or a phospholipase A 2-type activity. Sphingomyelin was employed as substrate for the venom hemolysin, and the organic and aqueous fractions of the reaction mixtures were analyzed by thin-layer chromatography. Analysis of the organic fraction revealed a phosphate-containing product with the solubility and chromatographic characteristics of N-acylsphingosine phosphate (ceramide phosphate), and analysis of the aqueous fraction demonstrated the presence of choline. The isolation and identification of these products indicate that the sphingomyelin of the red cell membrane is hydrolyzed by a sphingomyelinase D-type activity expressed by the partially purified venom hemolysin. A close correspondence between the hemolytic and sphingomyelinase D activities was observed when the partially purified hemolysin was further characterized in polyacrylamide gel electrophoresis at pH 8.3 and pH 4.9. The hemolytic and sphingomyelinase activities were coincident within the electrophoretic pattern at both pHs. The results presented demonstrate conclusively a direct lytic action of brown recluse venom upon red blood cells and report for the first time the presence of sphingomyelinase D in spider venom.

Phosphates in red cells of two lungfish: the South American, Lepidosiren paradoxa, and the African, Protopterus aethiopicus

Measurements were made of water-soluble phosphates in red blood cells of two lungfish, the South American, Lepidosiren paradoxa, and the African, Protopterus aethiopicus. Lepidosiren was studied on the research vessel Alpha Helix on the Amazon River and P. aethiopicus was obtained at Lake Victoria, Kenya. The red cells of both species contained moderate amounts of ATP and GTP, with the former predominant. Also present were higher concentrations of the uridine mononucleotides, UDP and UTP, than had been seen in other kinds of red cells. Of special interest was the discovery in red cells of both fish of a previously unreported inositol diphosphate. No other inositol polyphosphates were found. The red cells of P. aethiopicus had two phosphate compounds (one an adenine nucleotide), which are still unidentified

Phosphates in red cells of two South American osteoglossids: Arapaima gigas and Osteoglossum bicirrhosum

Water-soluble phosphates of red blood cells of the South American osteoglossid fish Arapaima gigas were measured by ion-exchange column chromatography. Major phosphate components were ATP and GTP with the former predominant. Of special interest was the presence of a large pool of inositol pentaphosphate, hitherto seen only in red cells of birds and turtles. A close relative of Arapaima, another South American osteoglossid, Osteoglossum bicirrhosum, had no inositol polyphosphates in its red cells. This study stemmed from work on the research vessel Alpha Helix on the Amazon River.

32P TRACER STUDIES ON THE METHODS OF INCREASING THE EFFICIENCY OF CITRATE SOLUBLE AND INSOLUBLE PHOSPHATES FOR RICE ON ACID SOILS

Studies with 32P-tagged mono-, di-, or tricalcium phosphate (MCP, DCP, TCP) showed that on application to moist aerobic acid soils, a considerable proportion of the applied phosphates even from DCP and TCP were converted to Al-P and/or Fe-P due to gradual dissolution by the H+ ions of the system, resulting in increased Olsen or Bray P. Dry matter production of rice and utilization of applied P was found to be higher when the P carriers were applied to moist acid soils 2 weeks before flooding than when these were applied at flooding. The reasons for this differential response to time of application have been explained. There appears scope for making the citrate soluble and insoluble phosphates as efficident as water soluble phosphates for growing rice on acid soils by their application to the moist soil 2 weeks before flooding.

Incorporation of N-acetyl-D-glucosamine from UDP-N-acetyl-D-glucosamine by isolated membranes of Bacillus subtilis. Identification of undecaprenyl poly(N-acetylglucosaminyl pyrophosphate).

Membrane isolated from Bacillus subtilis strain 168 incorporated GlcNAc from UDP-GlcNAc directly onto undecaprenyl phosphate via transphosphorylation and subsequent transglucosylations. Chain lengths of 6, 4, and 1 units of GlcNAc were found. Approximately 80% of the isotope incorporated was extracted into chloroform:methanol (2:1 v/v), and could be distinguished from the undecaprenyl disaccharide cell wall intermediate by a different elution pattern on DEAE-cellulose (acetate form). The GlcNAc-lipid(s) were eluted from a similar column in chloroform:methanol:water (10:10:3, v/v) with 6 mM NH4COOH indicating a pyrophosphate linkage between the lipid and the GlcNAc. The GlcNAc-lipid(s) were not degraded by conditions which completely deacylated [32P]glyceryl phospholipids, but were rapidly hydrolyzed by mild acid treatment (0.005 N HCl, 90 degrees) with the release of oligosaccharide phosphate (typical of sugars linked to undecaprenyl pyrophosphate). Catalytic hydrogenation of the GlcNAc-lipid(s) resulted in the release of water-soluble sugar phosphate. Under these same conditions, undecaprenyl pyrophosphate and undecaprenyl disaccharide cell wall intermediate were similarly effected while [32P]glyceryl phospholipids remained intact. The formation of GlcNAc-lipid(s) in vitro was inhibited if membranes were prepared from cells previously treated with bacitracin. Thus, the GlcNAc-lipid(s) has the properties of undecaprenyl poly(N-acetylglucosaminyl pyrophosphate) and may represent a new synthetic role of the polyisoprenyl lipid in B. subtilis.

Evaluation of some sources of fertilizer phosphorus in two cycles of a paddy-wheat cropping sequence

SUMMARYA field experiment was made on Gurdaspur clay loam to assess the relative efficiency of various sources of fertilizer phosphorus for a paddy-wheat cropping sequence. Urea ammonium phosphate, superphosphate, a nitrophosphate fertilizer containing 30% water-soluble P and fertilizers containing 50 and 70% water-soluble P prepared from the nitrophosphate and superphosphate, dicalcium phosphate and rock phosphate were applied to the paddy crop at the rate of 0, 15, 30, 45 and 60 kg p2O6/ha and to wheat at double these rates.The cumulative yield data for paddy and wheat show a slight reduction in efficiency of phosphatio fertilizers, with the exception of dicalcium phosphate, as their watersoluble content decreased below 70%. In general, the relative efficiency of the less water-soluble sources and of rock phosphate was considerably higher for paddy than for wheat.The results show that the relative efficiency of low water-soluble phosphate fertilizers increased with time. The efficiency of phosphorus fertilizers declined with the rate of their application, more so in the case of higher water-soluble materials.

Cytidine Diphosphate Diglyceride of Bovine Brain. Positional Distribution of Fatty Acids and Analysis of Major Molecular Species

A method is described for the isolation of CDP-diglyceride from bovine brain. Yields of the product ranged from 9.2-15.5 mumol per kilogram of tissue, which corresponds to about 1% of the level of phosphatidic acid. Mild alkaline hydrolysis of the product gave three water-soluble phosphate esters which had the same electrophoretic mobilities as CMP, CDP-glycerol and glycerol 3-phosphate. The liponucleotide was quantitatively hydrolysed by CDP-diglyceride hydrolase from Escherichia coli to phosphatidic acid and CMP. No dCMP was recovered in enzymatic or alkaline hydrolysates and it is concluded there can be little or no dCDP-diglyceride in bovine brain. Brain CDP-diglyceride was similar to phosphatidylinositol in that in both lipids stearate was the major saturated fatty acid and arachidonate the most abundant unsaturated fatty acid. This differed significantly from the fatty acid patterns of other metabolically related phospholipids, phosphatidic acid and cardiolipin. Brain CDP-diglyceride was hydrolysed with phospholipase C from Clostridium welchii with the liberation of the diglyceride moiety in high yield. Treatment of the diglyceride with pancreatic lipase showed CDP-diglyceride with the asymmetric distribution of fatty acids characteristic of most mammalian phospholipids, saturated fatty acids being found mostly at position 1 and polyunsaturated fatty acids at position 2. The derived diglyceride acetates were separated into different molecular species by argentation thin-layer chromatography. These analyses showed that 1-stearoyl, 2-arachidonoyl was the major species of brain CDP-diglyceride.

Reaction of synthetic corundum with orthophosphoric acid

It was shown in an Investigation of mixtures of fine-ground electrofused corundum with orthophosphoric acid that at 20–25°C the degree of the reaction of the components is highest during the first four days. During this period only water-soluble acid aluminum phosphates are formed and a significant proportion of the orthophosphoric acid remains uncombined. The reaction rate is low and falls still further with an increase in the size of the corundum grains.

The formation of cyclic inositol 1,2-monophosphate, inositol 1-phosphate, and glucose 6-phosphate by brain preparations stimulated with deoxycholate and calcium: A gas chromatographic study

A gas chromatographic method has been developed for the separation and isolation of water-soluble phosphates as trimethylsilyl ethers. With this method cyclic inositol 1,2-monophosphate and inositol 1-phosphate, derived from endogenous phosphatidylinositol, have been shown to increase when a particulate portion of brain homogenate is stimulated with deoxycholate and Ca ++, confirming earlier observations of Lapetina and Michell (1). Concomitant with the appearance of inositol phosphates is the stimulated formation of glucose 6-phosphate in the whole homogenate. Although ATP can replace deoxycholate and Ca ++ in a dialyzed homogenate, glucose 6-phosphate apparently does not arise by any known metabolic pathway but from another unidentified source.

Effects of the amounts of phosphate fertilizer, and of the proportions of water-soluble phosphate in the fertilizers tested on the phosphorus nutrition of sorghum

SUMMARYA field experiment was made with sorghum hybrid CSH-1 during the rainy seasons of 1968 and 1969 on a sandy loam soil to study the influence of amounts and sources of phosphate fertilizer on phosphorus nutrition. Concentration of P in the crop and uptake increased with increasing amounts of phosphate and with increasing proportions of water-soluble P in triple superphosphate-dicalcium phosphate and triple superphosphate-rock phosphate mixtures. Relative effectiveness values were calculated from P uptake at harvest by a multiple regression method. For triple superphosphate, nitrophosphate (30% water soluble P), dicalcium phosphate and rock phosphate (from Jordan) the values were respectively, 100, 74, 61 and 22.

Umwandlung und Verfügbarkeit von Rohphosphat (Hyperphos) verglichen mit aufgeschlossenen Phosphaten unter kontrollierten Temperatur‐ und Feuchtebedingungen

Transformation and availability of rock phosphate (Hyperphos) compared with other fertilizer phosphates under controlled temperature and moisture conditions.In order to characterize the efficient portion of a fertilizer after transformation in soil, in samples of typical soils of Schleswig‐Holstein the easily mobilizable and the mobile P‐content were investigated with specific extractants.The transformation under controlled varied temperature and moisture conditions results first (besides in mobile phosphates) in easily mobilizable Ca‐phosphate. In acid soils this changes gradually into Al‐ and Fe‐phosphate the content of which decreases due to ageing (transformation into hardly mobilizable forms). The pH is the most important factor influencing the kind of transformation whereas high temperature and moisture content mainly increase speed and extent (high temperature accelerating especially the ageing process). The water soluble phosphate is less influenced by temperatur and moisture.The best efficiency of rock phosphates results from rapid transformation (promoted by low pH) into easily mobilizable soil phosphate followed by an ageing‐retarding constellation of factors such as low temperature and moisture.

Growth characters and yield of sorghum (Sorghum vulgare Pers.) as affected by contents of water–soluble P in triple superphosphate/dicalcium phosphate and triple superphosphate/rock phosphate mixtures

SUMMARYA field experiment was made during the rainy seasons of 1968 and 1969 on a sandy loam soil to study the response of Sorghum hybrid CSH–1 to phosphorus in relation to content of water-soluble phosphate in fertilizers. Triple superphosphate, dicalcium phosphate, rock phosphate (200-mesh), nitrophosphate and mixtures of triple superphosphate with dicalcium phosphate and rock phosphate in 3:1, 2:2 and 1:3 ratios were studied. The amounts of phosphorus applied were 30 and 60 kg P2O5/ha. Phosphorus application significantly affected dry matterproduced, plant height, days to 50% bloom, number of grains per ear, thousand-grain weight, stover and grain yield. Triple superphosphate and 3:1 triple superphosphate/dicalcium phosphate mixture were the most effective while rock phosphate and 1:3 triple superphosphate/rock phosphate mixture the least. Differences among sources were more marked in 1968 and at the higher amount i.e. 60 kg P2O5/ha. The response of sorghum to phosphorus was considerably lowered when the water solubility waa below 50% in triple superphosphate/dicalcium phosphate mixtures and below 75% in triple superphosphate/rock phosphate mixtures. Relative effectiveness was assessed by calculating ‘superphosphate equivalents’ and ‘multiple regressions’. Using superphosphate equivalents dicalcium phosphate, nitrophosphate and rock phosphates were 47–53%, 35–56% and 3–25%, respectively, as effective as triple superphosphate.

The structure of a glycerylphosphoryldiglucosyl diglyceride from the lipids of Acholeplasma laidlawii strain B.

1. The phosphatidylglucose structure proposed previously (Smith & Henrikson, 1965) for the glucose-containing phospholipid from Acholeplasma laidlawii is incorrect. 2. The structure now proposed is 3-(sn-glycerol-3-phosphoryl-6'-[O-alpha-d-glucopyranosyl-(1-->2)-O-alpha-d-glucopyranosyl])- sn-1,2-diglyceride, a new type of bacterial lipid. 3. Deacylation of the lipid gave a single water-soluble phosphate ester which could be distinguished on chromatography from synthetic samples of glucosylphosphorylglycerols. 4. Hydrolysis of the lipid with alkali gave a mixture of fatty acids, glycerol 2-phosphate, sn-glycerol 3-phosphate and O-alpha-d-glucopyranosyl-(1-->2)-O-alpha- d-glucopyranosyl-(1-->1)-d-glycerol. 5. The lipid was unaffected on incubation with phospholipases A, C and D. 6. Diglucosyl diglyceride was isolated after treatment of the lipid with 60% HF, establishing the location of the fatty acid residues. 7. Periodate oxidation studies showed that the sn-glycerol 3-phosphate was esterified to the 6-hydroxyl group of one of the glucose residues in diglucosyl diglyceride.

Wheat yield responses to various phosphate fertilizers in South Australia

A number of phosphate fertilizers were tested as a source of phosphorus for wheat during three seasons (1967-1969) in the cereal growing areas of South Australia. The fertilizers were standard superphosphate, double superphosphate, calcium phosphate, two forms of calcined phosphate, a mixture of calcined phosphate with superphosphate, and untreated rock phosphate. Nine experiments were sown on a range of soils (pH 6.2 8.4) during the three years.At equivalent rates of total phosphorus, the water-soluble phosphates (superphosphate, double superphosphate) and the readily hydrolysed calcium pyrophosphate gave similar responses in all experiments. Water insoluble calcined and rock phosphates gave lower yields than superphosphate and, in most cases, the yields were not significantly greater than with no fertilizer.Uptake of phosphorus in the grain paralleled the grain yield differences between the fertilizers.

Water-soluble phosphates in horse granulocytes and lympocytes

1. 1. A study was made of the water-soluble phosphate compounds of combined leukocytes and purified lymphocytes and garnulocytes from fresh horse blood collected in heparin. 2. 2. The lymphocytes and granulocytes were each isolated in large quantities and high purity, after centrifugal separation from erythrocytes and platelets, by using columns of special glass beads. 3. 3. The phosphate compounds, which include nucleotides and glucolytic intermediates, were isolated from trichloroacetic acid extracts of the cells by chromatographic procedures in Dowex 1 (formate) columns and identified by a variety of chemical and enzymatic methods.

The effect of imbibition and vernalization on the uptake and distribution of 32P-orthophosphate in the grains and grain parts of a spring and winter wheat

32P-orthophosphate was taken up by both Marquis and Rideau wheat grains during imbibition at 24 °C and converted mainly into water-soluble organic phosphates and to a much smaller extent into the nucleic acid, phospholipid, and phosphoprotein components of both the whole grains and grain parts. Whereas the 32P incorporation into the nucleic acids and phospholipids of Rideau grains increased during the chilling period (0–5 weeks), the incorporation of 32P into these fractions was relatively low in Marquis grains and only slightly affected by cold treatment. The results suggest that the cold treatment results in overall stimulation of turnover or synthesis of nucleic acids, phosphoproteins, and membrane phospholipids in Rideau grains. The 32P incorporation data also indicate that the scutellum of both varieties acts as a "sink" for the phosphate metabolites from the other grain parts.

Relationship between phosphates and alkaline phosphatase of Anabaena flos-aquae in continuous culture.

Anabaena flos-aquae was grown in chemostats with phosphate-limiting growth and dilution rate of 0.015–0.03 h-1. The yields of cells were dependent on dilution rate and a two-fold increase obtained by growth in the presence of 15 mM KNO3. Alkaline phosphatase activity varied 20-fold, lowest activity with excess phosphate light-limited cells and the highest activity with cells grown in the presence of 15 mM KNO3. There was no correlation between hot water soluble phosphate of cells and alkaline phosphatase activity.

The permeability of the mitochondrial inner membrane to sucrose

The permeability of the inner membrane to sucrose in fresh and aged mitochrondria is described. Theoretical aspects of the use of the distribution of sucrose as a reference for movements of ions and water are discussed. Experimental findings support the following conclusions: 1. 1. When mitochondria are exposed to labelled sucrose for less than 5 min, penetration across the inner membrane is not significant; and the [ 14C]sucrose space is a reliable estimate of the volume external ot this membrane. The size of the inner, ion-containing compartment can then be estimated by different (total water minus sucrose-accessible water). 2. 2. With longer incubations, sucrose gains entrance into the inner compartment. Under these conditions, the sucrose-free space may continue to have meaning as a virtual space in which the endogeneous solutes are isotonic to the external medium. Changes in this virtual space remain useful for evaluating movements of ions and water across the inner membrane. 3. 3. Entrance of sucrose, and also of Cl −, into the inner compartment during aging was found to be closely linked to reciprocal losses of K + and water-soluble phosphate compounds.

Metabolism of phospholipids by spermatozoa and seminal plasma.

1. The hydrolysis of added (32)P-labelled phospholipids by whole ram and bull semen and the separated spermatozoal and plasma components was examined. 2. The ethanolamine phosphoglycerides were rapidly attacked by washed spermatozoa, forming predominantly glycerylphosphorylethanolamine, but with whole semen and seminal plasma a lysophosphatidylethanolamine was also detected. 3. The hydrolysis of lecithin by spermatozoa and plasma was very slow, and glycerylphosphorylcholine was the sole product detected. 4. Ram testicular spermatozoa were comparatively inactive in metabolizing both phospholipids, but ampulla contents showed the same activity as ejaculated semen. 5. Phosphatidylinositol was metabolized by spermatozoa obtained from any portion of the ram reproductive tract and also by seminal plasma. With testicular components, ampulla contents and washed ejaculated spermatozoa, inositol monophosphate, an unidentified phosphate ester and inorganic phosphate were the main products. In contrast, with whole semen and seminal plasma, glycerylphosphorylinositol was the predominant water-soluble phosphate ester. 6. Accessory-gland secretion obtained from vasectomized rams showed a pronounced phospholipase A activity towards ethanolamine phosphoglyceride. 7. On aerobic incubation of whole ram semen there was a decrease in the concentration of all phospholipid classes, although cardiolipin showed the greatest percentage decrease. In the choline phosphoglyceride fraction, this loss was confined to the plasmalogen component. This breakdown of phospholipids was decreased considerably when the spermatozoa were washed, and was not observed when whole bull semen was incubated under similar conditions.