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1.
The kinetics of sodium dependency of GABA uptake by satellite glial cells was studied in bullfrog sympathetic ganglia. GABA uptake followed simple Michaelis-Menten kinetics at all sodium concentrations tested. Increasing external sodium concentration increased bothK m andV max for GABA uptake, with an increase in theV max/K m ratio. The initial rate of uptake as a function of the sodium concentration exhibited sigmoid shape at 100 M GABA. Hill number was estimated to be 2.0. Removal of external potassium ion or 10 M ouabain reduced GABA uptake time-dependently. The effect of ouabain was potentiated by 100 M veratrine. These results suggest that at least two sodium ions are involved with the transport of one GABA molecule and that sodium concentration gradient across the plasma membrane is the main driving force for the transport of GABA. The essential sodium gradient may be maintained by Na+, K+-ATPase acting as an ion pump.  相似文献   

2.
Summary Sodium and chloride influxes across the nonshort-circuited isolated skin ofRana esculenta were measured at widely varying external ionic concentrations.The curve describing sodium transport has two Michaelis-Menten components linked at an inflection point occurring at an external sodium concentration of about 7 meq. Chloride transport can also be represented by two saturating components. A possible explanation of these kinetics is discussed.At sodium concentrations lower than 4 meq it is possible to define a component of the sodium transport mechanism as having a high affinity for sodium and which is independent of the nature of the external anion. A high affinity for chloride of the chloride transport system functioning at low external concentrations is also found but is significantly different from that of sodium. These systems show the physiological characteristics of the countertransports (Na ext + /H int + ; Cl ext /HCO 3int ) functioning at low external concentrations.At external concentrations higher than 4 meq a low affinity transporting system in which chloride and sodium are linked superimpose on the high affinity components.The physiological significance of these results is discussed.  相似文献   

3.
Asymmetric displacement currents, Ig, were measured in squid axons at different hydrostatic pressures, P, up to 60 MPa. Potassium and sodium currents were abolished by intracellular Cs+ and TEA+, by extracellular Tetrodotoxin (TTX), and by Na+ substitution with Tris+. The time course of Ig became progressively slower with increasing pressure, and the amplitude decreased. With appropriate scaling in time and amplitude, Ig records at any given P could be made to superimpose very well with those obtained at atmospheric pressure. The same scaling factors yielded a good superposition of all records obtained for voltage steps to membrane potentials in the range-30 to +42 mV. The ratio between the amplitude and time factors was larger than unity and increased with P, indicating a progressive decrease (up to 35% at 60 MPa) of the total charge displaced, Q, with no significant change in its voltage dependence. The time-scaling factor increased exponentially with P, as expected if all the steps involved in the opening of a sodium channel, and producing a major charge redistribution, have the same activation volume, V g 17 cm3/mol. This value is roughly one-half of that characterizing the pressure dependence of sodium current activation, suggesting that some late, rate-limiting step in the opening of sodium channels has a large activation volume without being accompanied by an easily detected charge movement.Part of the decrease of Q with pressure could be attributed to an increase in sodium inactivation. However, we cannot exclude the possibility that there is a reversible reduction in the number of fast activating sodium channels, similar to the phenomenon that has been reported to occur at low temperatures (Matteson and Armstrong 1982).  相似文献   

4.
Summary To assess the active components of sodium flux across toad bladder as a function of transepithelial potential, unidirectional sodium fluxes between identical media were measured before and after adding sufficient ouabain (1.89×10–3 m) to eliminate active transport, while clamping transepithelial potential to 0, 100 or 150 mV. Evidence was adduced that ouabain does not alter passive fluxes, and that fluxes remain constant if ouabain is not added. Hence, the ouabain-inhibitable fluxes represent fluxes through the active path. Results were analyzed by a set of equations, previously shown to describe adequately passive fluxes under electrical gradients in this tissue, here modified by the insertion ofE, the potential at which bidirectional sodium fluxes ( E and E ) through the active pathway are equal. According to these equations, E and E are the logarithmic mean of bidirectional fluxes through the active path at any potential, and the flux ratio in this path is modified by a constant factorQ ia, which represents the ratio of the bulk diffusion coefficient to the tracer diffusion coefficient in this pathway. The data are shown to conform closely to these equations.Q ia averages 2.54. Hence, serosal-to-mucosal flux vanishes rapidly as potential falls belowE. MeanE in these experiments was 158±1 mV. Thus, linear dependence of net flux in both active and passive pathways on potential is present, even though the sodium fluxes in both paths fail to conform to the Ussing flux ratio equation.Q i p<1 in the passive path (qualitatively similar to exchange diffusion) andQ ia>1 in the active path (as in single file pore diffusion). Both of these features tend to reduce the change in serosal-to-mucosal sodium flux induced by depolarization from spontaneous potential to zero potential (short-circuiting).  相似文献   

5.
Summary The possibility that sodium from the serosal bathing medium back-diffuses into the active sodium transport pool within the mucosal epithelial cell of the isolated toad bladder was examined by determining the effect on the metabolism of the tissue of removing sodium from the serosal medium. It was expected that if recycling of serosal sodium did occur through the active transepithelial transport pathway of the isolated toad bladder, removal of sodium from the serosal medium would reduce the rate of CO2 production by the tissue and enhance the stoichiometric ratio of sodium ions transported across the bladder per molecule of sodium transport dependent CO2 produced simultaneously by the bladder (J Na/J CO 2). The data revealed no significant change in this ratio (17.19 with serosal sodium and 16.13 after replacing serosal sodium with choline). Further, when transepithelial sodium transport was inhibited (a) by adding amiloride to the mucosal medium, or (b) by removing sodium from the mucosal medium, subsequent removal of sodium from the serosal medium, or (c) addition of ouabain failed to depress the basal rate of CO2 production by the bladder [(a) rate of basal, nontransport related, CO2 production (J CO2 b ) equals 1.54±0.52 with serosal sodium and 1.54±0.37 without serosal sodium; (b)J CO2 b equals 2.18±0.21 with serosal sodium and 2.09±0.21 without serosal sodium; (c) 1.14±0.26 without ouabain and 1.13±0.25 with ouabain; unite ofJ CO2 b are nmoles mg d.w.–1 min–1]. The results support the hypothesis that little, if any, recycling of serosal sodium occurs in the toad bladder.  相似文献   

6.
The feasibility of arrowroot (Marantha arundinacea) starch for alkaline protease production using an alkalophilic Bacillus lentus isolate was evaluated in batch fermentations in shake flasks and in a bioreactor under a range of conditions. A new arrowroot starch-casein medium (pH 10.2) was formulated having a composition (%, w/v): arrowroot starch 1, casein 1, sodium succinate 0.25, NH4Cl 0.05, NaCl 0.05, KH2PO4 0.04, K2HPO4 0.03, MgCl2 0.01, yeast extract 0.01 and Na2CO3 1.05. The isolate produced a maximum protease yield (6754.7 U ml–1) in this medium when grown for 72 h at 250 rev/min and 37 °C. Scaling-up studies in a bioreactor showed a 5-fold increase in alkaline protease yields (31899 U ml–1) at a lower production time of 45 h, aeration of 1 v/v/m and agitation of 400 rev/min at 37 °C.  相似文献   

7.
Summary Holthuisana transversa maintains sodium balance at very low external concentrations (7–25 M). This is achieved by the high affinity of the uptake mechanism (K m=0.18 mM), a low rate of sodium loss (0.17 mol g–1h–1) and a large increase in the rate of sodium uptake after depletion. The site of sodium absorption is the gills and extraction of sodium from respired water by sodium-depleted crabs can be as high as 28%. Crabs tolerate direct transfer to 80% sea-water and, following acclimation, the sodium and calcium levels of the haemolymph are maintained above those of the medium while the level of magnesium is significantly lower.  相似文献   

8.
The 17 base pair operator O R 3 oligonucleotide, which is the preferential binding site for the Cro repressor of phage , was studied by two-dimensional NMR spectroscopy. A sequential assignment procedure based on two-dimensional Nuclear Overhauser Effect (NOESY) and scalar coupling correlated (COSY) NMR spectroscopy, together with the knowledge of the oligodesoxynucleotide sequence, made it possible to assign the non-exhangeable base protons and the H1 and the H2-H2 sugar protons of the O R 3 operator DNA. The pattern of the observed NOE connectivities is consistent with a right-handed helical DNA structure. The base and sugar proton assignments provide the necessary information for further studies of the O R 3 operator — Cro repressor interaction.Abbreviations COSY correlated spectroscopy - FID free induction decay - NOE nuclear Overhauser effect - NOESY nuclear Overhauser effect spectroscopy - RD relaxation delay - TSP sodium 3-trimethylsilyl-(2,2,3,3-2H4)propionate - EDTA sodium ethylendiamine tetraacetate  相似文献   

9.
The chloroplast coupling factor (CF1) was dissociated into subunits by the freezing-thawing procedure in the presence of 0.5 M NaBr and the subunit was purified by ion-exchange chromatography on a DEAE-cellulose column. The subunit did not catalyze ATP hydrolysis either in the presence or in the absence of reagents known to activate Mg2+-dependent ATPase activity of CF1. However, it manifested appreciable adenylate kinase-like and ATP-ADP -phosphate exchange activities. The adenylate kinase-like activity only slightly depended on Mg2+ ions. Ethanol, and especially diadenosine pentaphosphate, inhibited the reaction effectively. In contrast, the ATP-ADP exchange activity was Mg2+-dependent. Ethanol and diadenosine pentaphosphate were poor inhibitors. Sulfite, the CF1-ATPase activator, and quercetin, its inhibitor, had a minor effect on catalytic activity of the subunit.Abbreviations CF chloroplast coupling factor 1 - RBP carboxylase-ribulose-1,5-bisphosphate carboxylase - TLC thin layer chromatography - MES morpholinoethane sulfonic acid - PMSF phenylmethylsulfonyl fluoride - AP5A diadenosine pentaphosphate, P1, P5-bis(5-adenosyl)pentaphosphate - DCCD N1N-dicyclohexylcarbodiimide - SDS sodium dodecylsulfate - PAAG polyacrylamide gel  相似文献   

10.
Summary Pulsed field gradients were incorporated into the HCACO experiment for acquiring spectra on isotopically enriched protein samples dissolved in H2O. Excellent water suppression and spectral quality were achieved using the modified pulse sequence (gd-HCACO), as demonstrated for a 13C-/15N-labeled sample of the SH2 domain from the hematopoietic cellular kinase dissolved in 90% H2O/10% D2O. Strong correlations for all residues were observed in the gd-HCACO spectrum, even for residues having -protons resonating exactly at the H2O frequency. The HCACO-TOCSY experiment was modified to correlate intraresidue 13C (rather than 1H), carbonyl (13C), and aliphatic side-chain protons [(H)CACO-TOCSY]. Pulsed field gradients were also incorporated into the (H)CACO-TOCSY experiment for water suppression.  相似文献   

11.
Spin-State-Selective Excitation (S3E), which forexample selectively excites amide proton resonances corresponding toexclusively either the or the spin state of the covalentlybound 15N atom is employed for E.COSY-type extraction ofheteronuclear J coupling constants. Instead of having one spectrum with twopeaks (corresponding to the or spin state of15N), S3E generates two spectra, each with onlyone peak for each 15N nucleus. These two spectra are generatedfrom the same data set, so that there is no reduction in sensitivitycompared to conventional 1JNH-resolved methods.Another interesting feature in comparison with conventional methods is that1JNH can be suppressed during the evolutionperiod, meaning that no heteronuclear multiplet structure is visible in the1 frequency dimension. The S3E pulsesequence element is combined with NOESY for measurement of3JN-H and JN-Hcoupling constants in either a hetero- or a homonuclear correlated version.Experimental confirmation is obtained using the protein RAP 17-;97(N-terminal domain of 2-macroglobulin ReceptorAssociated Protein).  相似文献   

12.
A novel halophilic strain that could carry out heterotrophic nitrification and aerobic denitrification was isolated and named as Halomonas campisalis ha3. It removed inorganic nitrogen compounds (e.g. NO3 ?, NO2 ? and NH4 +) simultaneously, and grew well in the medium containing up to 20 % (w/v) NaCl. PCR revealed four genes in the genome of ha3 related to aerobic denitrification: napA, nirS, norB and nosZ. The optimal conditions for aerobic denitrification were pH 9.0, at 37 °C, with 4 % (w/v) NaCl and sodium succinate as carbon source. The nitrogen removal rate was 87.5 mg NO3 ?–N l?1 h?1. Therefore, this strain is a potential aerobic denitrifier for the treatment of saline wastewater.  相似文献   

13.
Two respiratory-deficient nuclear petites, FY23Δpet191 and FY23Δcox5a, of the yeast Saccharomyces cerevisiae were generated using polymerase-chain-reaction-mediated gene disruption, and their respective ethanol tolerance and productivity assessed and compared to those of the parental grande, FY23WT, and a mitochondrial petite, FY23ρ0. Batch culture studies demonstrated that the parental strain was the most tolerant to exogenously added ethanol with an inhibition constant. K i, of 2.3% (w/v) and a specific rate of ethanol production, q p, of 0.90 g ethanol g dry cells−1 h−1. FY23ρ0 was the most sensitive to ethanol, exhibiting a K i of 1.71% (w/v) and q p of 0.87 g ethanol g dry cells−1 h−1. Analyses of the ethanol tolerance of the nuclear petites demonstrate that functional mitochondria are essential for maintaining tolerance to the toxin with the 100% respiratory-deficient nuclear petite, FY23Δpet191, having a K i of 2.14% (w/v) and the 85% respiratory-deficient FY23Δcox5a, having a K i of 1.94% (w/v). The retention of ethanol tolerance in the nuclear petites as compared to that of FY23ρ0 is mirrored by the ethanol productivities of these nuclear mutants, being respectively 43% and 30% higher than that of the respiratory-sufficient parent strain. This demonstrates that, because of their respiratory deficiency, the nuclear petites are not subject to the Pasteur effect and so exhibit higher rates of fermentation. Received: 22 September 1997 / Accepted: 7 December 1997  相似文献   

14.
The chick blastoderm at the stage of late gastrula is a flat disc formed by three cell layers and exhibiting epithelial properties. Blastoderms were cultured in miniature chambers and their electrophysiological characteristics were determined under Ussing conditions.Under open-circuit condition and identical physiological solutions on both sides, spontaneous transblastodermal potential difference (V oc) of –7.5±3.3 mV (ventral side positive) was measured. Under short-circuit condition (transblastodermal V = 0 mV), the blastoderm generated short-circuit current (I sc) of 21±8 A/cm2, which was entirely dependent on extracellular sodium, sensitive to ouabain applied ventrally and independent of extracellular chloride. The net transblastodermal Na+ flux fully accounted for the measured I sc, both under control conditions and with ouabain. The total transblastodermal resistance (R tot) was 390±125 cm2.Frequently, the V oc, I sc and R tot showed spontaneous oscillations with a period of 4–5 min. Removal of endoderm and mesoderm did not significantly affect the electrical properties, indicating that the electrogenic sodium transport is generated by the ectoderm.The V oc and I sc measured in the area pellucida (–1.3±0.8 mV, 9.3±4.4 A/cm2) and extraembryonic area opaca (–7.8±1.1 mV, 31.2±12.7 A/cm2) were significantly different. Such a heterogeneous distribution of electrical properties can explain the presence in the blastoderm of extracellular electrical currents found by using a vibrating probe.This work was supported by the Swiss National Research Foundation (grant. 3.418-0.86 to P.K.) and by Roche Research Foundation (grant. to U.K.). We thank Drs. E. Raddatz and Y. de Ribaupierre for helpful discussions.  相似文献   

15.
The lobster (Homarus americanus) hepato-pancreatic epithelial baso-lateral cell membrane possesses three transport proteins that transfer calcium between the cytoplasm and hemolymph: an ATP-dependent calcium ATPase, a sodium-calcium exchanger, and a verapamil-sensitive cation channel. We used standard centrifugation methods to prepare purified hepato-pancreatic baso-lateral membrane vesicles and a rapid filtration procedure to investigate whether 65Zn2+ transfer across this epithelial cell border occurs by any of these previously described transporters for calcium. Baso-lateral membrane vesicles were osmotically reactive and exhibited a time course of uptake that was linear for 10–15 s and approached equilibrium by 120 s. In the absence of sodium, 65Zn2+ influx was a hyperbolic function of external zinc concentration and followed the Michaelis-Menten equation for carrier transport. This carrier transport was stimulated by the addition of 150 M ATP (increase in Km and Jmax) and inhibited by the simultaneous presence of 150 mol l–1 ATP+250 mol l–1 vanadate (decrease in both Km and Jmax). In the absence of ATP, 65Zn2+ influx was a sigmoidal function of preloaded vesicular sodium concentration (0, 5, 10, 20, 30, 45, and 75 mmol l–1) and exhibited a Hill Coefficient of 4.03±1.14, consistent with the exchange of 3 Na+/1Zn2+. Using Dixon analysis, calcium was shown to be a competitive inhibitor of baso-lateral membrane vesicle 65Zn2+ influx by both the ATP-dependent (Ki=205 nmol l–1 Ca2+) and sodium-dependent (Ki=2.47 mol l–1 Ca2+) transport processes. These results suggest that zinc transport across the lobster hepato-pancreatic baso-lateral membrane largely occurred by the ATP-dependent calcium ATPase and sodium-calcium exchanger carrier proteins.Communicated by: I.D. Hume  相似文献   

16.
The sodium ion gradient and the membrane potential were found to be the driving forces of sulfate accumulation in the marine sulfate reducer Desulfovibrio salexigens. The protonmotive force of –158 mV, determined by means of radiolabelled membrane-permeant probes, consisted of a membrane potential of –140 mV and a pH gradient (inside alkaline) of 0.3 at neutral pHout. The sodium ion gradient, as measured with silicone oil centrifugation and atomic absorption spectroscopy, was eightfold ([Na+]out/[Na+]in) at an external Na+ concentration of 320 mM. The resulting sodium ionmotive force was –194 mV and enabled D. salexigens to accumulate sulfate 20000-fold at low external sulfate concentrations (<0.1 M). Under these conditions high sulfate accumulation occurred electrogenically in symport with three sodium ions (assuming equilibrium with the sodium ion-motive force). With increasing external sulfate concentrations sulfate accumulation decreased sharply, and a second, low-accumulating system symported sulfate electroneutrally with two sodium ions. The sodium-ion gradient was built up by electrogenic Na+/H+ antiport. This was demonstrated by (i) measuring proton translocation upon sodium ion pulses, (ii) studying uptake of sodium salts in the presence or absence of the electrical membrane potential, and (iii) the inhibitory effect of the Na+/H+ antiport inhibitor propylbenzilylcholin-mustard HCl (PrBCM). With resting cells ATP synthesis was found after proton pulses (changing the pH by three units), but neither after pulses of 500 mM sodium ions, nor in the presence of the uncoupler tetrachorosalicylanilide (TCS). It is concluded that the energy metabolism of the marine strain D. salexigens is based primarily on the protonmotive force and a protontranslocating ATPase.Abbreviations MOPS morpholinopropanesulfonic acid - TCS tetrachlorosalicylanilide - PrBCM propylbenzilylcholin-mustard HCl - Tris tris(hydroxymethyl)aminomethane - TPP+ bromide tetraphenylphosphonium bromide  相似文献   

17.
The Mg-nucleoside triphosphatase activity associated with the inner envelope membrane of the pea chloroplast is comprised of at least two components, a major activity that is sensitive to vanadate and sodium fluoride and a minor insensitive activity. The vanadate/fluoride sensitive activity has been partially purified (about 35-fold) from Triton X-100 solubilized membranes by DEAE-Sephadex chromatography and sucrose density gradient centrifugation. The partially purified enzyme resembles the membrane-bound activity in requiring either Mg2+ or Mn2+, having a broad specificity for nucleoside triphosphates, having a Km for ATP of 0.18 millimolar, and being inhibited by N-ethylmaleimide, but insensitive to sodium azide and dicyclohexylcarbodiimide. The partially purified enzyme obtained after sucrose gradient centrifugation has a markedly increased sensitivity to inhibition by inorganic pyrophosphate compared with the less pure enzyme. Pyrophosphate is not a substrate of either the membrane-bound or partially purified enzyme.  相似文献   

18.
The gas phase holdup and mass transfer characteristics of carboxymethyl cellulose (CMC) solutions in a bubble column having a radial gas sparger have been determined and a new flow regime map has been proposed. The gas holdup increases with gas velocity in the bubbly flow regime, decreases in the churn-turbulent flow regime, and increases again in the slug flow regime. The volumetric mass transfer coefficient (k La) significantly decreases with increasing liquid viscosity. The gas holdup and k La values in the present bubble column of CMC solutions are found to be much higher than those in bubble columns or external-loop airlift columns with a plate-type sparger. The obtained gas phase holdup ( g) and k La data have been correlated with pertinent dimensionless groups in both the bubbly and the churn-turbulent flow regimes.List of Symbols a m–1 specific gas-liquid interfacial area per total volume - A d m2 cross-sectional area of downcomer - A r m2 cross-sectional area of riser - d b m individual bubble diameter - d vs m Sauter mean bubble diameter - D c m column diameter - D L m2/s oxygen diffusivity in the liquid - Fr Froude number, U g/(g Dc)1/2 - g m/s2 gravitational acceleration - G a Galileo number, gD c 3 2/2 app - H a m aerated liquid height - H c m unaerated liquid height - K Pa · sn fluid consistency index - k L a s–1 volumetric mass transfer coefficient - n flow behavior index - N i number of bubbles having diameter d bi - Sc Schmidt number, app/( D L) - Sh Sherwood number, k L a D c 2 /DL - U sg m/s superficial gas velocity - U gr m/s superficial riser gas velocity - V a m3 aerated liquid volume - V c m3 unaerated liquid volume - N/m surface tension of the liquid phase - g gas holdup - app Pa · s effective viscosity of non-Newtonian liquid - kg/m3 liquid density - ý s–1 shear rate - Pa shear stress  相似文献   

19.
Summary The relationships between intracellular sodium content, sodium transport and serum effects were investigated in human fibroblasts. In the cells with low intracellular sodium (Na iL /+ ;0.04 mol sodium/mg protein) serum stimulated the sodium-potassium pump as measured by ouabain-sensitive sodium efflux and rubidium influx and also exerted a transstimulation of ouabain-insensitive sodium transport resulting in net influx. In cells with high intracellular sodium (Na iH /+ ;0.42 mol sodium/mg protein) all aspects of sodium transport were increased compared to Na iL /+ cells. In these cells serum caused no change in sodium-potassium pump activity but significantly increased the ouabain-insensitive sodium fluxes resulting in net efflux. In Na iL /+ cells, serum promoted net sodium influx through an amiloride-sensitive pathway that was undetectable in the basal state. In Na iH /+ cells the serum-stimulated net efflux was amiloride sensitive but this pathway also contributed to a major portion of sodium transport in the basal state. This study demonstrated that sodium-potassium pump activity is directed by the supply of internal sodium and that serum can increase this supply by promoting net influx, and that serum-induced sodium transport can be modified by intracellular sodium content.  相似文献   

20.
The aim of the current study was to evaluate the cardioprotective ability of water (WE) and ethanolic (EE) papaya fruits extracts against cardiotoxicity induced by aflatoxin B1 (AFB1) in rats. Forty two female Sprague–Dawley rats were divided into six treatment groups and treated orally for 2 weeks as follow: control group, the group treated with WE (250 mg/kg b.w), the group treated with EE (250 mg/kg b.w), the group treated orally with AFB1 (17 μg/kg b.w) and the groups treated orally with AFB1 plus WE or EE. The results indicated that treatment with AFB1 resulted in oxidative stress in the heart manifested by the marked increase in cardiac malondialdehyde and calcium levels accompanied with a significant decrease in cardiac total antioxidant capacity. Serum nitric oxide and sodium levels, lactate dehydrogenase and creatine kinase isoenzyme activities were significantly increased, whereas, cardiac Na+/K+-ATPase activity and serum potassium were insignificantly affected. Supplementation with WE or EE effectively ameliorated most of the changes induced by AFB1. It could be concluded that both extracts attenuated the oxidative stress induced in heart tissue by AFB1 and WE was more pronounced due to the higher total phenolic contents than in the EE.  相似文献   

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