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1.
Summary A gel forms on the surface of nodal roots of corn. We have made observations relevant to parameters of its molecular size,
the nature of its interaction with clay, and the actual existence of it under normal root growth. In so doing we have anticipated
that it is similar to if not the same as the mucilagenous gel at the root soil interface, and that the knowledge gained would
be useful in understanding the processes operating at this interface.
We have found that gel is present all along a nodal root surface that had elongated into a tube filled with water saturated
sand. This observation emphasizes that gel is present under near natural conditions. We have estimated, using a modification
of the light scattering technique, that the exudate has a molecular weight of approximately 9 × 107. According to X-ray patterns of the c-axis of exudate films, it appears as if it is an amorphous material. IR spectra of
it revealed that it has a characteristic polysaccharide pattern. Addition of exudate to NH4-montmorillonite caused a large increase in the intensity of the first order c-axis X-ray diffraction peak and a very slight
increase in the calculated d-spacing. The results were interpreted as consistent with the idea that exudate addition to the
clay provided a media for better dispersal of the clay plates, however it was not intercalated into the clay lattice. These
observations suggest that the exudate may play an important function on the root-soil interface, however more work needs to
be done to understand its action at the root-soil interface.
Contribution from the Department of Agronomy, Purdue University, Lafayette, Indiana, U.S. Journal Paper No.3971 of the Purdue Agricultural Experiment Station. 相似文献
2.
The reversible delipidation of a solubilized sodium-plus-potassium ion-dependent adenosine triphosphatase from the salt gland of the spiny dogfish.
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P Ottolenghi 《The Biochemical journal》1975,151(1):61-66
A microsomal fraction rich in Na+, K+-ATPase (sodium-plus-potassium ion-dependent adenosine triphosphatase) and the corresponding K+-dependent p-nitrophenyl phosphatase from the rectal salt gland of the spiny dogfish was solubilized by treatment with deoxycholate at high ionic strength. On gel filtration through Sepharose 6B, the ATPase apoenzyme could be separated, in apparently soluble form, from the tissue-fraction phospholipids and was almost free of enzymic activity (2% of the p-nitrophenyl phosphatase activity and 0.2% of the ATPase activity being recovered). On mixing the apoenzyme with an activator consisting of cooked ox brain, a large proportion of the original enzymic activity was obtained. Specific activities of the re-activated enzyme were somewhat higher than in the material before gel filtration: values of 1300-1450 mumol and 250-290 mumol/h per mg of protein were obtained for the hydrolysis of ATP and of p-nitrophenyl phosphate respectively. The activity was inhibitible by ouabain. 相似文献
3.
Properties of a series of tegumental membrane-bound phosphohydrolase activities of Schistosoma mansoni. 总被引:2,自引:0,他引:2
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1. Incubation of Schistosoma mansoni for 5 min in a phosphate-buffered medium, pH 7.4, released tegumental material containing the following phosphohydrolase activities: alkaline phosphatase, 5'-nucleotidase, glycerol-2-phosphatase, glucose 6-phosphatase, phosphodiesterase and ATPase. 2. Maximum activity of these enzymes was measured at pH 9.5; however, the phosphodiesterase and ATPase activities were also appreciable at pH 7.0. 3. Solubilization of the released tegumental material in 1% Triton X-100 followed by gel filtration distinguished three peaks of enzyme activity: an ATPase (mol.wt. greater than 1000 000), a phosphodiesterase (mol.wt. 1 000 000) and an alkaline phosphomonoesterase with broad specificity (mol.wt. 232 000). 4. The ATPase activity was highly activated by 10 mM-Mg2+ or 1 mM-Ca2+ and was inhibited by chelating agents. Ouabain, Na+ and K+ had little effect on enzyme activity, whereas activity was increased by 50% in the presence of calmodulin. The phosphodiesterase activity was highest in the presence of 100 mM-Na+ or -K+, and 10 mM-Mg2+ or -Ca2+. Alkaline phosphatase activity was also stimulated by 100 mM-Na+ or -K+, and 10 mM-Mg2+; however Ca2+ inhibited at greater than 1 mM. 5. Surface iodination of parasites followed by detergent solubilization and gel filtration of the released tegumental membranes indicated that these enzymes were not accessible. A major surface component, apparent mol.wt. 80 000, was iodinated. 6. Rabbit anti-(mouse liver 5'-nucleotidase) antibodies did not inhibit the phosphohydrolase activities. However, an immunoglobulin G fraction from sera of mice chronically infected with S. mansoni partially inhibited alkaline phosphatase activity, but was without effect on the phosphodiesterase and ATPase activities. 7. The location of the enzymes in the double membrane of the tegument and their significance in host-parasite interactions is discussed. 相似文献
4.
Phosphatases in cucumber roots, whose production was inducedby Ca2$ deficiency, were characterized chromatographically usingATP, 2'(3')-AMP and p-nitrophenyl-phosphate (PNPP) as substrates.Ca2$ deficiency stimulated greater than 10-fold increases inthe activities with these substrates of the non-adsorbed fractionfrom a DEAE-cellulose column. Several fractions associated withthese phosphatase activities were eluted from the column withNaCl solution; their levels increased less with Ca2$ starvation.When the non-adsorbed fraction from Ca2$-straved roots was appliedto a Sephadex G-200 column, fractions associated with 2'(3')-AMPase(phosphatase I) and with both ATPase and PNPPase (phosphataseII) were separated. In the control roots, very weak activitiesof phosphatases I and II were observed at the same positionon the gel filtration. The phosphatase I isolated from boththe control and Ca2$-starved roots was extremely specific tonucleoside 2'(3')-monophosphates, whereas phosphatase II fromboth types of roots had a relatively broad substrate specificity.When phosphatase I from Ca2$-starved roots was stained with2'(3')-AMP in CaCl2 after polyacrylamide gel electrophoresis,a single band was obtained. Phosphatase I from control rootsalso showed a single band, with the same Rf value. PhosphataseII from both types of roots contained two isozyme bands whenthe activities were stained with either ATP or PNPP. These resultsindicate that Ca2$ starvation causes specific increases in thelevel of phosphatases I and II in cucumber roots. (Received October 28, 1981; Accepted January 19, 1982) 相似文献
5.
Purification and characterization of the proton translocating plasma membrane ATPase of red beet storage tissue 总被引:1,自引:0,他引:1
Plasma membranes were prepared from red beet (Beta vulgaris L.) storage tissue by partition in an aqueous two-phase system. A highly active proton-translocating ATPase was purified from these membranes by lysophosphatidylcholine extraction and glycerol density gradient centrifugation. The ATPase activity was inhibited by vanadate or dicyclohexyl carbodiimide, but was insensitive to azide, nitrate and molybdate at concentrations which inhibit the F1ATPase, the tonoplast ATPase, and acid phosphatase. Inhibition by vanadate was consistent with a non-competitive mechanism, with Ki = 10 microM. The Km for Mg-ATP was about 1 mM, magnesium ions were required, and the activity was stimulated by KCl and by lysophosphatidylcholine. The optimal pH was 6.5. The molecular mass by gel filtration in the presence of 2 g/liter octyl glucoside was 600 kDa, while dodecyl sulfate gel electrophoresis gave a polypeptide molecular mass of 100 kDa. After blotting onto nitrocellulose, the purified enzyme did not bind concanavalin A, although a concanavalin A-binding peptide of the plasma membrane runs to nearly the same position on the gel and showed some tendency to co-purify with the ATPase. Phospholipid vesicles into which the purified ATPase had been incorporated by the freeze-thaw technique showed vanadate-sensitive, ATP-dependent proton uptake. When the ATPase was reconstituted into lipid membranes at high protein to lipid ratios and incubated with ATP, two-dimensionally crystalline arrays of protein molecules were formed. 相似文献
6.
At least nine acid phosphatases and a distinct phytase are present in different cell fractions of germinating lettuce. The enzymes could be partially characterised using acrylamide gel electrophoresis. Phosphatase formation is only partially inhibited by cycloheximide. A new soluble ATPase arises between 24 and 48 hours of germination. Its formation is not inhibited by cycloheximide. Phosphatase activity in the particulate fraction of the cell can be liberated and activated by detergent or trypsin treatment. It is suggested that the newly formed soluble ATPase arises by release and activation of a particulate phosphatase. 相似文献
7.
The isoform spectra of alkaline and acid phosphatase, pyrophosphataes, and ATPase in periplasm of E. coli were studied using electrophoresis in polyacrylamide gel with subsequent development of zimograms directly in the gel. Wild strains and mutants on 4 regulatory genes of alkaline phosphatase were analyzed. Mutations in regulatory genes were shown to influence the amount of each of the 3 isoforms of alkaline phosphatase and also the spectra of other phosphohydrolases. 相似文献
8.
Wagner GJ 《Plant physiology》1981,68(2):499-503
The membrane of anthocyanin containing Hippeatrum petal vacuoles was examined for protein and enzyme content after purification by equilibrium density centrifugation. Light scattering, protein, and a Mg2+-dependent nucleotide specific ATPase were associated with membrane having a density of 1.08 to 1.12 grams per cubic centimeter. A small amount of acid phosphatase was also present in this region of the gradient, but this activity peaked at about 1.12 grams per cubic centimeter. A component of yeast tonoplast, α-mannosidase, was not significantly present. UDP-glucose, anthocyanidin-3-O-glucosyltransferase, thought to be a cytosol enzyme in Hippeastrum, was absent from tonoplast of vacuoles isolated by osmotic shock in 0.2 molar K2HPO4 or 0.35 molar mannitol. Vacuolar acid phosphatase was insensitive to ethylenediaminetetraacetate but was 80% inhibited by 10 millimolar KF, while ATPase was inactivated by 2 millimolar ethylenediaminetetraacetate and only 50% inhibited by 10 millimolar KF. Five major and about 9 minor polypeptides were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of membrane protein on 5 to 30 and 6 to 16% gradient gels. 相似文献
9.
Renal brush border membrane vesicles (BBMV) of the dog possess at least two ATPase activities. In the present study, we have examined the effect of pH, ions, and inhibitors on the activity of ATPase in BBMV. Two different sets of conditions were identified that produced stimulation of ATPase activity. A unique stimulation of BBMV ATPase activity occurred at acidic pH in the presence of 1 mM ZnCl2. In the absence of Zn2+, a second ATPase activity was stimulated by alkaline pH values with peak stimulation occurring between pH 8.5 and 9.0. The results suggest that the alkaline pH-stimulated hydrolysis of ATP probably represents the activity of BBMV alkaline phosphatase. The unique acidic pH + Zn2(+)-stimulated ATPase activity must represent the activity of a second protein other than the alkaline phosphatase, since purified alkaline phosphatase did not show this activity. The biochemical identity and physiological function of this renal BBMV ATPase activity remain to be determined, but it may be an ecto-ATPase. 相似文献
10.
In order to examine acid phosphatase (EC 3.1.3.2) and ATPase (EC 3.6.1.3) activities of baker's yeast (pH optimum 3.5) a protoplast-secreted enzyme preparation was purified and some physical and chemical properties were studied. Three protein fractions containing ATPase and acid phosphatase activities, in the same ratio as the initial preparation, were separated by ion-exchange chromatography. The first fraction which had the highest protein content yielded a homogeneous preparation after Sepharose 4B chromatography and was used in further studies. An attempt to estimate molecular weight of this protein was made. Attempts to separate acid phosphatase and ATPase activities by ion-exchange chromatography, gel filtration, isoelectric focusing and sucrose density gradient centrifugation have been unsuccessful. Both activities behaved the same way to heat and urea denaturation. These results suggest that the two activities are associated with the same protein molecule. 相似文献
11.
Root growth and nitrate uptake by wheat (Triticum aestivum L.) following wetting of dry surface soil 总被引:1,自引:0,他引:1
Brady D.J.; Wenzel C.L.; Fillery I.R.P.; Gregory P.J. 《Journal of experimental botany》1995,46(5):557-564
The nitrate uptake capacity of surface roots of spring wheat(Triticum aestivum L. cv. Kulin) was investigated followingwetting of dry surface soil. Plants were grown to stem elongationstage with adequate watering at depth while the surface soilwas allowed to dry. Eight weeks after sowing, water or a 15N-nitratesolution was added to the surface soil to simulate rainfall.Root growth and nitrate uptake were measured up to 4 d afterwetting on plants with unconfined nodal root growth and on plantswith the majority of nodal roots confined within small vials.Prior to wetting, plants from both nodal treatments had seminalroots with collapsed cortices along the upper 10 cm and manyshort, viable lateral roots. Nodal roots, where present, wereonly a few cm long and unbranched. Only plants with unconfinednodal roots were able to take up nitrate within the 24 h beforeany new root growth. By 2 d after wetting there was significantgrowth of the seminal lateral roots, and rapid growth and branchingof nodal roots. From 2 d after wetting, plants with confinednodal roots also took up nitrate, presumably due to the growthof the seminal lateral roots. Hence it appears as though thenodal roots in the unconfined treatment could immediately takeup nitrate, but the seminal roots required new lateral rootgrowth to become active in nitrate uptake. The plants with confinednodal roots had a lower nitrate uptake than those with unconfinednodal roots 4 d after wetting, indicating that the seminal rootsystem was not able to compensate for lack of nodal roots. Insufficientnitrate was taken up after 4 d, by plants from either nodalroot treatment, to increase the shoot N concentration significantly. Key words: Triticum aestivum, nitrate uptake, drought, seminal roots, nodal roots 相似文献
12.
The ATPase activity of the plasma membrane-enriched fractionwas severely inhibited by withdrawal of Ca2+ from the mediumfor 5 days, although the root system appeared to be unaffectedto visual inspection. Partially lipid-depleted ATPases withsimilar ratios of phospholipid to protein were prepared fromthe plasma membrane-enriched fraction of cucumber roots culturedwith control medium and one lacking Ca2+, and their propertieswere compared. SDS disc polyacrylamide gel electrophoresis showedthat the polypeptide components were essentially similar betweencontrol and Ca2+-starved roots. Partially lipid-depleted ATPasereassociated with asolectin, the lecithin from soybean, showedtypical characteristics of plasma membrane type ATPase; pH optimumat 6.5, high specificity for ATP as substrate and strong inhibitionby vanadate but not nitrate. The activity of reassociated ATPaseobtained from the control roots was apparently higher than theactivity obtained from Ca2+-starved roots. The amount of asolectinrequired for maximum activation of the partially lipid-depletedATPase prepared from control roots was much lower than thatprepared from Ca2+-starved roots. Reassociation of partiallylipid-depleted ATPase with asolectin produced higher ATPaseactivity than that with individual phospholipids. The activationof partially lipid-depleted ATPase prepared from control rootswith asolectin was not inhibited by addition of a sample preparedfrom Ca2+-starved roots. Thus, a decrease in the functionalassociation of ATPase with phospholipids might be one of thephysiological injuries in root cell membranes of cucumber causedby Ca2+ starvation.
1Permanent address: Department of Horticulture, College of Agriculture,Chonnam National University, Chonnam 500, Korea. (Received February 23, 1988; Accepted August 18, 1988) 相似文献
13.
ATPase (ATP phosphohydrolase, EC 3.6.1.3) activity was shown in the soluble fraction of rat liver micochondria. Two molecular forms (ATPase 1 and 2) were isolated. ATPase 1 has already been studied. The present paper deals with the purification method of ATPase 2 which was achieved by the following steps: (NH4)2SO4 precipitation. DEAE-cellulose chromatography, hydroxyapatite chromatography, Sephadex G100 filtration and AMP-Sepharose affinity chromatography. The purified protein was characterized by bidimensional polyacrylamide gel electrophoresis. Molecular weight evaluated by SDS-polyacrylamide gel electrophoresis and Sephadex G100 gel filtration was found to be 61 500 +/- 3000. 相似文献
14.
Observations were made at 2 or 4 wk intervals from December to harvest on all stages of Heterodera avenae in winter oats growing on infested land. Second-stage larvae were present in all soil samples except on 5 and 20 July. Invasion and development of larvae was slow during winter. The nodal and seminal roots of winter oats were both heavily invaded by the nematode; larvae which invaded seminal roots tended to become male whereas those in nodal roots tended to become female. There was a small second invasion in August. Females were first observed on the roots of winter oats on 17 May, 214 days after the crop was sown and 62 days after the first fourth-stage larva was observed. The nodal roots of spring barley contained few H. avenae larvae whereas these roots were heavily invaded in winter wheat and oats. In spring barley the nodal roots were developing in June and July when few second-stage larvae were in the soil whereas in winter oats and wheat the nodal roots were growing rapidly in April when larvae were most numerous, and so were heavily invaded. 相似文献
15.
Differential effects of lipid depletion on membrane sodium-plus-potassium ion-dependent adenosine triphosphatase and potassium ion-dependent phosphatase.
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The phospholipid-dependence of the (Na-++K-+)-dependent ATPase (adenosine triphosphatase) (EC 3.6.1.3) and associated K-+-dependent phosphatase activity (EC 3.6.1.7) have been compared. Unlike the (Na-++K-+)-dependent ATPase activities, the K-+-dependent phosphatase activities of a number of different preparations were not closely correlated with their total phospholipid contents. After partial lipid depletion with a single extraction in Lubrol W the residual ATPase and phosphatase activities were correlated, but their magnitudes were quite different: on average only about 5% of the former remained compared with 50% of the latter. A similar differential effect on these activities was found after extraction with deoxycholate. In contrast with the ATPase, consistent restoration of the phosphatase activity of Lubrol-extracted enzymes by added exogenous phospholipids was not observed. We conclude that, although the K-+-dependent phosphatase may be lipid-dependent, the lipid requirement must be different from that of the complete ATPase system, and this difference should help investigations of their relationship. 相似文献
16.
Effect of vanadate, molybdate, and azide on membrane-associated ATPase and soluble phosphatase activities of corn roots 总被引:18,自引:41,他引:18
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The effects of vanadate, molybdate, and azide on ATP phosphohydrolase (ATPase) and acid phosphatase activities of plasma membrane, mitochondrial, and soluble supernatant fractions from corn (Zea mays L. WF9 × MO17) roots were investigated. Azide (0.1-10 millimolar) was a selective inhibitor of pH 9.0-ATPase activity of the mitochondrial fraction, while molybdate (0.01-1.0 millimolar) was a relatively selective inhibitor of acid phosphatase activity in the supernatant fraction. The pH 6.4-ATPase activity of the plasma membrane fraction was inhibited by vanadate (10-500 micromolar), but vanadate, at similar concentrations, also inhibited acid phosphatase activity. This result was confirmed for oat (Avena sativa L.) root and coleoptile tissues. While vanadate does not appear to be a selective inhibitor, it can be used in combination with molybdate and azide to distinguish the plasma membrane ATPase from mitochondrial ATPase or supernatant acid phosphatase.
Vanadate appeared to be a noncompetitive inhibitor of the plasma membrane ATPase, and its effectiveness was increased by K+. K+-stimulated ATPase activity was inhibited by 50% at about 21 micromolar vanadate. The rate of K+ transport in excised corn root segments was inhibited by 66% by 500 micromolar vanadate.
相似文献17.
A J Burgess I Matthews E A Grimes A M Mata F M Munkonge A G Lee J M East 《Biochimica et biophysica acta》1991,1064(1):139-147
m-Maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) was used to cross-link the protein components of rabbit skeletal muscle sarcoplasmic reticulum. Analysis of cross-linked material by SDS-polyacrylamide gel electrophoresis showed that both the (Ca(2+)-Mg2+)-ATPase and the 53 kDa glycoprotein could be cross-linked, since the amount of protein at the locations on the gel corresponding to uncross-linked material was reduced in the presence of 1.0 mM MBS. Cross-linked products of 130 kDa, 200-260 kDa and approx. 300 kDa were identified. Probing the cross-linked products with monoclonal antibodies against ATPase, 53 kDa glycoprotein and calsequestrin revealed no cross-linked products containing the ATPase and either calsequestrin or the 53 kDa glycoprotein over the range of molecular weights examined here. Possible interactions between the ATPase and calsequestrin or the 53 kDa glycoprotein were also investigated by studying the ATPase activity for the purified ATPase and for the ATPase in sarcoplasmic reticulum vesicles made permeable to Ca2+ with A23187. Effects of Ca2+ and ATP on the two systems were indistinguishable, providing no evidence for a major modulatory role of calsequestrin or the 53 kDa glycoprotein on the ATPase. 相似文献
18.
Purification and Properties of the Plasma Membrane H-Translocating Adenosine Triphosphatase of Phaseolus mungo L. Roots 总被引:5,自引:4,他引:1
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Kasamo K 《Plant physiology》1986,80(4):818-824
The plasma membrane ATPase of mung bean (Phaseolus mungo L.) roots has been solubilized with a two-step procedure using the anionic detergent, deoxycholate (DOC) and the zwitterionic detergent, zwittergent 3-14 as follows: (a) loosely bound membrane proteins are removed by treatment with 0.1% DOC; (b) The ATPase is solubilized with 0.1% zwittergent in the presence of 1% DOC; (c) the solubilized material is further purified by centrifugation through a glycerol gradient (45-70%). Typically, about 10% of the ATPase activity is recovered, and the specific activity increases about 11-fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that the peak fraction from the glycerol gradient contains three major polypeptides of Mr = 105,000, 67,000, and 57,000 daltons. The properties of the purified ATPase are essentially the same as those of membrane-bound ATPase, with respect to pH optimum, substrate specificity, inhibitor sensitivity, and ion stimulation. 相似文献
19.
Adaptation of H+-pumping and plasma membrane H+ ATPase activity in proteoid roots of white lupin under phosphate deficiency 总被引:14,自引:0,他引:14
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White lupin (Lupinus albus) is able to adapt to phosphorus deficiency by producing proteoid roots that release a huge amount of organic acids, resulting in mobilization of sparingly soluble soil phosphate in rhizosphere. The mechanisms responsible for the release of organic acids by proteoid root cells, especially the trans-membrane transport processes, have not been elucidated. Because of high cytosolic pH, the release of undissociated organic acids is not probable. In the present study, we focused on H+ export by plasma membrane H+ ATPase in active proteoid roots. In vivo, rhizosphere acidification of active proteoid roots was vanadate sensitive. Plasma membranes were isolated from proteoid roots and lateral roots from P-deficient and -sufficient plants. In vitro, in comparison with two types of lateral roots and proteoid roots of P-sufficient plants, the following increase of the various parameters was induced in active proteoid roots of P-deficient plants: (a) hydrolytic ATPase activity, (b) Vmax and Km, (c) H+ ATPase enzyme concentration of plasma membrane, (d) H+-pumping activity, (e) pH gradient across the membrane of plasmalemma vesicles, and (f) passive H+ permeability of plasma membrane. In addition, lower vanadate sensitivity and more acidic pH optimum were determined for plasma membrane ATPase of active proteoid roots. Our data support the hypothesis that in active proteoid root cells, H+ and organic anions are exported separately, and that modification of plasma membrane H+ ATPase is essential for enhanced rhizosphere acidification by active proteoid roots. 相似文献
20.
Acrosomal ATPase in starfish and bivalve mollusk spermatozoa 总被引:1,自引:0,他引:1
ATPase activity was found in acrosomes of starfish and bivalve mollusk spermatozoa, using a cytochemical method with electron microscopy. The activity was located in central material of the starfish acrosome and in material lining the acrosomal membrane of the Mytilus acrosome, as well as in the basal part of the starfish acrosome. The ATPase activity in the former material was preferably activated by Ca2+, while that in the starfish basal material was preferably activated by Mg2+. Both types of activity persisted during and after the acrosome reaction. ATPase activity was also observed in the region of the axial filament complex of the flagella, in centrioles and in a basal matrix. ATPase in the acrosome also hydrolysed other nucleoside triphosphates. However, there was no detectable phosphatase activity, and little pyrophosphatase or 5′-nucleotidase activity. Evidence was obtained that adenylate kinase may be included in the acrosome. A possible role of the ATPase activity in the acrosome reaction is discussed. 相似文献