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1.
The effect of pH of electrolyte solution on the interfacial tension of lipid membrane formed of phosphatidylcholine (PC, lecithin)–phosphatidylserine (PS) system was studied. In this article, three models describing the H+ and OH ions adsorption in the bilayer lipid surface are presented. In Model I and Model II, the surface is continuous with uniformly distributed functional groups constituting the centres of H+ and OH ions adsorption while in the other the surface is built of lipid molecules, free or with attached H+ and OH ions. In these models contribution of the individual lipid molecule forms to interfacial tension of the bilayer were assumed to be additive. In Model III the adsorption of the H+ and OH ions at the PC–PS bilayer surface was described in terms of the Gibbs isotherm. Theoretical equations are derived to describe this dependence in the whole pH range.  相似文献   

2.
N. Mano  A. Kuhn   《Biosensors & bioelectronics》2001,16(9-12):653-660
We report about the electrocatalytic properties of electrodes modified by adsorption of nitro-fluorenone derivatives. The stable, adherent monolayer of these catalyst precursors can be transformed electrochemically into the corresponding hydroxylamine compounds (R-NO2+4e+4H+R-NHOH+H2O). The completely reversible two electron oxidation of the hydroxylamine leads to the nitroso compounds (R-NHOHR-NO+2e+2H+) that exhibit high catalytic activity in the electrooxidation of NADH at low overpotentials (−30 mV vs. Ag/AgCl) and therefore constitute a new family of efficient redox mediators for biosensor applications. A significant increase in catalytic activity (up to 500%) is observed after addition of calcium ions to the electrolyte. This is explained by a specific and bridging complexation between the coenzyme's phosphate groups and a carboxyl group present in the catalyst molecule. The interaction favours the contact between NADH and the surface confined catalyst, leading to a higher electron transfer efficiency. This interaction can be used in an approach of molecular level design for controlled monolayer deposition of catalyst, Ca2+, NAD+ and enzyme. A very simple and inexpensive modification scheme, essentially based on electrostatic attraction, leads to electrodes that can be employed as reagentless biosensors for the electrochemical detection of common and commercially interesting analytes like glucose.  相似文献   

3.
磷饥饿条件下番茄幼苗的H+分泌速率明显提高。质膜质子泵专一性抑制剂钒酸盐能显著抑制番茄幼苗的H+分泌,也能显著抑制其Pi吸收。此结果表明,磷饥饿时番茄幼苗Pi吸收速率的变化与H+分泌速率的变化之间可能具有一定的相关性,并进一步暗示质膜H+-ATPase可能参与其中。本文结果还表明,Pi/H+的准量关系约为1:1。  相似文献   

4.
Robert L. Heath 《BBA》1972,256(3):645-655
ATP formation by isolated chloroplasts is due to the proton gradient phenomena, according to Mitchell. The number of protons moved per electron pair transported and per photon absorbed is related to the number of protons required to produce each ATP. Thus, a critical test of the Mitchell hypothesis is the quantum yield of H+ transport. Bromocresol purple, a pH indicator, can be used to measure the pH external to isolated chloroplasts accurately and rapidly. The action spectrum (with pycocyanine as the electron acceptor) appears to be that of a System I-linked reaction (high above 700 nm). The quantum yield has been calculated to be 3.5 ± 0.1 H+/hv from 640 nm to 690 nm and 6.7 ± 0.4 H+/hv above 700 nm. The action spectrum of the efflux of H+ occurring in the dark, which is usually identified as being equivalent to the steady-state influx, has the same shape as that of the influx. The quantum yield, however, is reduced by 0.5. Therefore, Photosystem II seems to affect both the initial influx and dark efflux. The H+/photon and H+/e2 for the initial influx are too high for the Mitchell hypothesis. Only the H+ efflux in the dark from 640–690 nm has an H+/hv of 1.6 which agrees with the theory of Mitchell.  相似文献   

5.
Tonoplast H+-ATPase and H+-pyrophosphatase (H+-PPase) were previously characterized in Acer pseudoplatanus cells (A. Pugin et al., Plant Sci., 73 (1991) 23–34; A. Fraichard et al., Plant Physiol. Biochem., 31 (1993) 349–359). The present study concerns the relationships between these two enzymes in vitro. ATP and PPi hydrolysis were additive and the inhibition of one did not affect the activity of the second one. ATP and PPi H+-transports were also additive. The H+ -PPase inhibition did not change ATP-dependent H+-transport but H+-ATPase inhibition inhibited the PPi dependent H+-transport. Because H+-PPase was reported to transport H+ and K+ into the vacuole (Davies et al., Proc. Natl. Acad. Sci. USA, 89 (1992) 11701–11705), these results led us to suggest that the inhibition of the H+-ATPase activity could modify the H+/K+ stoichiometry for the benefit of K+-transport.  相似文献   

6.
Light-dependent Ca2+ efflux via the Ca2+/H+ antiport in the photosynthetic purple sulfur bacterium Chromatium vinosum was inhibited by three phenothiazines: chlorpromazine; trifluoperazine and phenothiazine. The inhibitors had no effect on Ca2+ uptake by C. vinosum in the dark nor any effect on the light-dependent efflux of either Na+ or Tl+ catalyzed, respectively, by the C. vinosum Na+/H+ or K+/H+ antiports. Ruthenium red and LaCl3, neither of which inhibited light-dependent Ca2+ efflux in C. vinosum, markedly inhibited Ca2+ uptake in the dark by C. vinosum cells. Ruthenium red had no effect on the uptake of either Na+or the K+ analog T1+ by C. vinosum cells in the dark. These results have been interpreted in terms of two separate Ca2+ transport systems in C. vinosum: (i) a phenothiazine-sensitive and ruthenium red, La3+-insensitive Ca2+/H+ antiport responsible for Ca2+ efflux in the light; and (ii) a ruthenium red and La3+-sensitive but phenothiazine-insensitive Ca2+ uptake system.  相似文献   

7.
The Mg2+-dependent H+-ATPase activity of a sealed microsomal vesicle fraction isolated from corn (Zea mays L.) roots appears to be controlled by a phosphorylation-dephosphorylation cycle. Phosphorylation of the microsomal fraction is carried out by a Ca2+/calmodulin (CaM)-stimulated process. The H+-ATPase activity decreases with increasing phosphorylation of the membranes and becomes only slightly uncoupled by ionophores and less inhibited by dicyclohexylcarbodiimide (DCCD), diethylstilbestrol (DES), NO3 and vanadate. The inhibitory effect of phosphorylation is greater on the NO3-sensitive H+-ATPase activity than on the vanadate-sensitive activity. Restoration of H+-ATPase activity is achieved by allowing the phosphorylated membranes to dephosphorylate either in the absence or presence of exogenous alkaline phosphatase. Moreover, the presence of fluphenazine during the Ca2+/CaM-stimulated treatment inhibits membrane phosphorylation and protects the H+-ATPase activity from inhibition.  相似文献   

8.
We examined the Na+/H+ exchanger message in isolated perfused rabbit hearts using Northern blot analysis with cDNA encoding for the rabbit cardiac Na+/H+ exchanger. A cDNA probe from the coding region of the rabbit myocardial Na+/H+ exchanger hybridized to mRNA of 5 kb under high stringency, and to a second 3.8 kb mRNA species under low stringency. When Northern blots were re-probed with a section of the 3′-untranslated region of the cDNA, the 5 kb message was apparent while the smaller 3.8 kb message was not. If isolated working rabbit hearts were subjected to ischemia we observed increases in the 3.8 kb message. Overall, the results show that a 3.8 kb mRNA product, which is homologous to the amiloride sensitive Na+/H+ exchanger, exists in the myocardium and increases during ischemia in the myocardium.  相似文献   

9.
采用外源一氧化氮(NO)供体硝普钠(SNP)研究了NO对盐胁迫下小麦(Triticum aestivum L.)幼苗耐盐性的影响.结果表明,0.1 mmol/L SNP处理显著缓解了1 50 mmol/L NaCl胁迫对小麦幼苗生长的抑制效应,包括水分丧失以及叶绿素降解,从而提高了小麦幼苗的耐盐性.进一步结合1 mg/mL血红蛋白处理则显著逆转了SNP诱导的上述效应;利用亚硝酸钠和铁氰化钾作为对照也证实了NO对小麦幼苗耐盐性的专一性调节作用,并可能与NO对小麦幼苗根部质膜H -ATPase和焦磷酸酶活性诱导有关.此外,尽管NO显著提高了盐胁迫下小麦幼苗根部细胞质膜H -ATPase和焦磷酸酶的ATP水解活性,但是对跨膜H 转运则没有明显影响.应用外源CaSO4和EGTA处理也证实,Ca2 可能在NO诱导的质膜H -ATPase和焦磷酸酶活性的提高过程中起信号作用.另外,分析盐胁迫下小麦幼苗根部Na 和K 含量的变化也发现,NO对Na 含量没有明显影响,但是却显著提高了K 水平和K /Na 比,这可能也是NO提高小麦幼苗耐盐性的原因之一.  相似文献   

10.
A N Malviya  P Anglard 《FEBS letters》1986,200(2):265-270
Transformed 3T3/10T1/2 cultured cells incubated with ferricyanide caused a decrease of 2 mM EDTA extractable cytosolic protein kinase C activity in 2 min, whereas 5 or 20 min ferricyanide treatment reverted the enzyme activity to that observed without ferricyanide. The ferricyanide effect in 2 min was abolished by amiloride and sustained by ouabain. Thus, deactivation-activation of cytosolic protein kinase C is attributed to an unknown signal generation during H+ accumulation coupled with the Na+/H+ exchange phase. In this mechanism the priming event concerns the transmembrane redox process shedding H+ into the cell interior while impermeant ferricyanide acts as a unique electron acceptor.  相似文献   

11.
J.Michael Gould  S. Izawa 《BBA》1974,333(3):509-524
1. By using dibromothymoquinone as the electron acceptor, it is possible to isolate functionally that segment of the chloroplast electron transport chain which includes only Photosystem II and only one of the two energy conservation sites coupled to the complete chain (Coupling Site II, observed P/e2 = 0.3–0.4). A light-dependent, reversible proton translocation reaction is associated with the electron transport pathway: H2O → Photosystem II → dibromothymoquinone. We have studied the characteristics of this proton uptake reaction and its relationship to the electron transport and ATP formation associated with Coupling Site II.

2. The initial phase of H+ uptake, analyzed by a flash-yield technique, exhibits linear kinetics (0–3 s) with no sign of transient phenomena such as the very rapid initial uptake (“pH gush”) encountered in the overall Hill reaction with methylviologen. Thus the initial rate of H+ uptake obtained by the flash-yield method is in good agreement with the initial rate estimated from a pH change tracing obtained under continuous illumination.

3. Dibromothymoquinone reduction, observed as O2 evolution by a similar flash-yield technique, is also linear for at least the first 5 s, the rate of O2 evolution agreeing well with the steady-state rate observed under continuous illumination.

4. Such measurements of the initial rates of O2 evolution and H+ uptake yield an H+/e ratio close to 0.5 for the Photosystem II partial reaction regardless of pH from 6 to 8. (Parallel experiments for the methylviologen Hill reaction yield an H+/e ratio of 1.7 at pH 7.6.)

5. When dibromothymoquinone is being reduced, concurrent phosphorylation (or arsenylation) markedly lowers the extent of H+ uptake (by 40–60%). These data, unlike earlier data obtained using the overall Hill reaction, lend themselves to an unequivocal interpretation since phosphorylation does not alter the rate of electron transport in the Photosystem II partial reaction. ADP, Pi and hexokinase, when added individually, have no effect on proton uptake in this system.

6. The involvement of a proton uptake reaction with an H+/e ratio of 0.5 in the Photosystem II partial reaction H2O → Photosystem II → dibromothymoquinone strongly suggests that at least 50% of the protons produced by the oxidation of water are released to the inside of the thylakoid, thereby leading to an internal acidification. It is pointed out that the observed efficiencies for ATP formation (P/e2) and proton uptake (H+/e) associated with Coupling Site II can be most easily explained by the chemiosmotic hypothesis of energy coupling.  相似文献   


12.
The release of protons from intact cells of Rhodopseudomonas capsulata after either 4μs flashes or during brief periods of continuous illumination has been measured with the indicator, cresol red. The half-time for H+-release after a flash was 35 ms and the extent, 1H+ per 134 bacteriochlorophyll. Myxothiazol completely inhibited the flash-induced H+-release and antimycin A reduced it by 37%. The proton-releasing reaction is discussed with reference to the protonmotive Q-cycle. During continuous illumination the rapid phase of H+ release is followed by a lag and then by another period of acidification, suggesting that other protolytic reactions may be in operation.  相似文献   

13.
Tea plants grow in acidic soil, but to date, their intrinsic mechanisms of acidic stress tolerance have not been elucidated. Here, we assessed the tea plant response to growth on NHt4 nutrient media having different p H and iron levels. When grown in standard NHt4 nutrient solution(iron insufficient, 0.35 mg Là1 Fe2t), tea roots exhibited significantly lower nitrogen accumulation, plasma membrane Ht-ATPase activity, and protein levels; net Htefflux was lower at pH 4.0 and 5.0 than at pH 6.0. Addition of30 mg Là1 Fe2t(iron sufficient, mimicking normal soil Fe2tconcentrations) to the NHt4 nutrient solution led to more efficient iron plaque formation on roots and increased root plasma membrane Ht-ATPase levels and activities at p H 4.0 eland 5.0, compared to the p H 6.0 condition. Furthermore,plants grown at pH 4.0 and 5.0, with sufficient iron,exhibited significantly higher nitrogen accumulation than those grown at pH 6.0. Together, these results support the hypothesis that efficient iron plaque formation, on tea roots, is important for acidic stress tolerance. Furthermore,our findings establish that efficient iron plaque formation is linked to increased levels and activities of the tea root plasma membrane Ht-ATPase, under low pH conditions.  相似文献   

14.
The period (∼3-5 min) of the ultradian rhythm of the lateral leaflet movement of Desmodium motorium is strongly lengthened (≤30-40%) by the K+ channel blocker tetraethylammoniumchloride (20, 30, and 40 mM) and vanadate (0.5 and 1 mM), which is an effective inhibitor of the plasma membrane-bound H+ pump. The alkali ions K+, Na+, Rb+, and Cs+ (10-40 mM) shorten the period only slightly (≤ 10-15%). Li+ (5-30 mM), however, increases the period of the leaflet rhythm drastically (≤80%). We concluded that the plasmalemma-H+-ATP-ase-driven K+ transport through K+ channels is an essential component of the ultradian oscillator of Desmodium, as has been proposed for the circadian oscillator.  相似文献   

15.
L. Sas  Z. Rengel  C. Tang 《Plant science》2001,160(6):61-1198
In symbiotically-grown legumes, rhizosphere acidification may be caused by a high cation/anion uptake ratio and the excretion of organic acids, the relative importance of the two processes depending on the phosphorus nutritional status of the plants. The present study examined the effect of P deficiency on extrusions of H+ and organic acid anions (OA) in relation to uptake of excess cations in N2-fixing white lupin (cv. Kiev Mutant). Plants were grown for 49 days in nutrient solutions treated with 1, 5 or 25 mmol P m−3 Na2HPO4 in a phytotron room. The increased formation of cluster roots occurred prior to a decrease in plant growth in response to P deficiency. The number of cluster roots was negatively correlated with tissue P concentrations below 2.0 g kg−1 in shoots and 3 g kg−1 in roots. Cluster roots generally had higher concentrations of Mg, Ca, N, Cu, Fe, and Mn but lower concentrations of K than non-cluster roots. Extrusion of protons and OA (90% citrate and 10% malate) from roots was highly dependent on P supply. The amounts of H+ extruded per unit root biomass decreased with time during the experiment. On the equimolar basis, H+ extrusion by P-deficient plants (grown at 1 and 5 mmol P m−3) were, on average, 2–3-fold greater than OA exudation. The excess cation content in plants was generally the highest at 1 mmol P m−3 and decreased with increasing P supply. The ratio of H+ release to excess cation uptake increased with decreasing P supply. The results suggest that increased exudation of OA due to P deficiency is associated with H+ extrusion but contributes only a part of total acidification.  相似文献   

16.
《植物生态学报》2017,41(4):489
Aims Elaeagnus angustifolia is one of the most salt-tolerant species. The objective of this study was to understand the mechanisms of ion transporation in E. angustifolia exposed to different salt concentrations through manipulations of K+/Na+ homeostasis.
Methods Seedlings of two variants of the species, Yinchuan provenance (YC, salt-sensitive type) and the Alaer provenance (ALE, salt-tolerant type), were treated with three different NaCl application modes, and the ion fluxes in the apical regions were measured using non-invasive micro-test technology (NMT). In mode 1, Na+ and K+ fluxes were measured after 150 mmol·L-1 NaCl stress lasted for 24 h. In mode 2, K+ and H+ fluxes were quantified with a transient stimulation of NaCl solution. In mode 3, Amiloride (Na+/H+ antiporters inhibitor) and tetraethylammonium (TEA, K+ channel inhibitor) was used to treat apical regions of E. angustifolia seedlings after NaCl stress for 24 h, respectively.
Important findings Under NaCl stress for 24 h, net effluxes of Na+ and K+ were increased significantly. The net Na+ effluxes of YC provenance seedlings (720 pmol·cm-2•s-1) were lower than that of ALE provenance (912 pmol·cm-2·s-1), but the net K+ efflux was higher in YC provenance. Under the instantaneous NaCl stimulation, net K+ efflux was remarkably increased, with the net K+ efflux of YC provenance always higher than that of ALE provenance. Interestingly, H+ at the apical regions was found from influx to efflux, with the net H+ efflux of ALE provenance greater than that of the YC provenance. Under the NaCl and NaCl + Amiloride treatment, the net Na+ efflux of ALE provenance seedlings was higher than that of YC provenance, while the net K+ efflux was less in ALE provenance seedlings. On the other hand, the differences in net Na+ and K+ effluxes were insignificant between the two provenances under the control group and NaCl + TEA treatment. In conclusion, NaCl stress caused Na+ accumulation and K+ outflows of E. angustifolia seedlings; The E. angustifolia seedlings utilize Na+/H+ antiporters to reduce Na+ accumulation by excretion; and the maintenance of K+/Na+ homeostasis in salt-tolerant E. angustifolia provenance seedlings roots accounted for a greater Na+ extrusion and a lower K+ efflux under NaCl stress. Results from this study provide a theoretical basis for further exploring salt-tolerant E. angustifolia germplasm resource.  相似文献   

17.
Auxin is unique among plant hormones in that its function requires polarized transport across plant cells. A chemiosmotic model was proposed to explain how polar auxin transport is derived by the H+ gradient across the plasma membrane (PM) established by PM H+-adenosine triphosphatases (ATPases). However, a classical genetic approach by mutations in PM H+-ATPase members did not result in the ablation of polar auxin distribution, possibly due to functional redundancy in this gene family. To confirm the crucial role of PM H+-ATPases in the polar auxin transport model, we employed a chemical genetic approach. Through a chemical screen, we identified protonstatin-1 (PS-1), a selective small-molecule inhibitor of PM H+-ATPase activity that inhibits auxin transport. Assays with transgenic plants and yeast strains showed that the activity of PM H+-ATPases affects auxin uptake as well as acropetal and basipetal polar auxin transport. We propose that PS-1 can be used as a tool to interrogate the function of PM H+-ATPases. Our results support the chemiosmotic model in which PM H+-ATPase itself plays a fundamental role in polar auxin transport.  相似文献   

18.
Although considerably more oxidation-resistant than other P-type ATPases, the yeast PMA1 H+-ATPase of Saccharomyces cerevisiae SY4 secretory vesicles was inactivated by H2O2, Fe2+, Fe- and Cu-Fenton reagents. Inactivation by Fe2+ required the presence of oxygen and hence involved auto-oxidation of Fe2+ to Fe3+. The highest Fe2- (100 μM) and H2O2 (100 mM) concentrations used produced about the same effect. Inactivation by the Fenton reagent depended more on Fe2+ content than on H2O2 concentration, occurred only when Fe2+ was added to the vesicles first and was only slightly reduced by scavengers (mannitol, Tris, NaN3, DMSO) and by chelators (EDTA, EGTA, DTPA, BPDs, bipyridine, 1, 10-phenanthroline). Inactivation by Fe- and Cu- Fenton reagent was the same; the identical inactivation pattern found for both reagents under anaerobic conditions showed that both reagents act via OH·. The lipid peroxidation blocker BHT prevented Fenton-induced rise in lipid peroxidation in both whole cells and in isolated membrane lipids but did not protect the H+-ATPase in secretory vesicles against inactivation. ATP partially protected the enzyme against peroxide and the Fenton reagent in a way resembling the protection it afforded against SH-specific agents. The results indicate that Fe2+ and the Fenton reagent act via metal-catalyzed oxidation at specific metal-binding sites, very probably SH-containing amino acid residues. Deferrioxamine, which prevents the redox cycling of Fe2+, blocked H+-ATPase inactivation by Fe2+ and the Fenton reagent but not that caused by H2O2, which therefore seems to involve a direct non-radical attack. Fe-Fenton reagent caused fragmentation of the H+-ATPase molecule, which, in Western blots, did not give rise to defined fragments bands but merely to smears.  相似文献   

19.
以甜瓜品种‘羊角酥瓜’为试材,利用人工气候室控制环境条件(昼/夜25/18 ℃),研究盐胁迫条件下外源褪黑素(MT)和Ca2+对甜瓜幼苗根系和叶片中Cl-、Na+、K+、Mg2+、Ca2+离子含量,Na+/K+、 Na+/Ca2+、Na+/Mg2+值,以及H+-ATP酶活性、渗透调节物质积累和细胞膜质过氧化的影响.结果表明: 与对照相比,盐胁迫处理显著抑制甜瓜幼苗生长,增加根系和叶片中Cl-、Na+含量,降低K+、Mg2+、Ca2+含量.盐胁迫下,喷施外源MT或Ca2+处理均可以显著降低甜瓜根系和叶片中Cl-、Na+含量,提高K+、Mg2+、Ca2+含量,植株体内Na+/K+、Na+/Ca2+和 Na+/Mg2+值下降;同时也提高了根系和叶片H+-ATP酶活性及叶片渗透调节物质的含量,降低盐胁迫对细胞膜的伤害,表现在甜瓜叶片相对电导率和丙二醛含量降低.总之,在盐胁迫条件下,外源MT、Ca2+单独和复配处理均可通过提高H+-ATP酶活性来降低盐害离子的含量,改善甜瓜幼苗中的离子平衡,同时增加渗透调节物质的含量,降低膜质过氧化水平,从而增强其对盐胁迫的适应性,其中MT和Ca2+复配处理时的效果更好.复配外施 MT 和Ca2+在诱导甜瓜幼苗提高耐盐方面具有协同增效作用.  相似文献   

20.
以甜瓜品种‘羊角酥瓜’为试材,利用人工气候室控制环境条件(昼/夜25/18 ℃),研究盐胁迫条件下外源褪黑素(MT)和Ca2+对甜瓜幼苗根系和叶片中Cl-、Na+、K+、Mg2+、Ca2+离子含量,Na+/K+、 Na+/Ca2+、Na+/Mg2+值,以及H+-ATP酶活性、渗透调节物质积累和细胞膜质过氧化的影响.结果表明: 与对照相比,盐胁迫处理显著抑制甜瓜幼苗生长,增加根系和叶片中Cl-、Na+含量,降低K+、Mg2+、Ca2+含量.盐胁迫下,喷施外源MT或Ca2+处理均可以显著降低甜瓜根系和叶片中Cl-、Na+含量,提高K+、Mg2+、Ca2+含量,植株体内Na+/K+、Na+/Ca2+和 Na+/Mg2+值下降;同时也提高了根系和叶片H+-ATP酶活性及叶片渗透调节物质的含量,降低盐胁迫对细胞膜的伤害,表现在甜瓜叶片相对电导率和丙二醛含量降低.总之,在盐胁迫条件下,外源MT、Ca2+单独和复配处理均可通过提高H+-ATP酶活性来降低盐害离子的含量,改善甜瓜幼苗中的离子平衡,同时增加渗透调节物质的含量,降低膜质过氧化水平,从而增强其对盐胁迫的适应性,其中MT和Ca2+复配处理时的效果更好.复配外施 MT 和Ca2+在诱导甜瓜幼苗提高耐盐方面具有协同增效作用.  相似文献   

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