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71.
Göran Wingstrand 《Physiologia plantarum》1985,64(2):223-229
Membrane-bound MgATPase activity from roots of young sugar beet ( Beta vulgaris L. cv. Monohill) was investigated in a membrane fraction purified by partition in an aqueous polymer two-phase system. After two steps of "washing" with fresh bottom phase (rich in dextran), the polyethylene glycol rich top phase (U3 ) was practically free of mitochondrial membranes (cytochrome oxidase), and the remaining MgATPase activity showed high substrate specificity for ATP. An optimum for the MgATPase activity was found at pH 7. The activation by Na+ or K+ was strongest on the acid side without any observable shift in pH optimum. Oligomycin had no effect, but vanadate strongly inhibited the U3 MgATPase and the K+ activation was lost. The complex activation pattern achieved by varying the Na+ /K+ ratio at constant total concentration was interpreted as a synergistic (Na+ + K+ )-activation. The U3 fraction MgATP-ase activity showed a 4-fold increase in the presence of 0.01% Triton X-100 implying that the MgATPase activity is located in vesicles of which 75% or more are sealed with the ATP binding site on the inside. Comparison with the properties of plasma membrane. ATPases from other plants indicated that the U3 fraction MgATPase was mainly of plasma membrane origin. 相似文献
72.
The colon of L. maderae consists of a single columnar epithelium covered with a cuticle and of a musculo-connective sheath. The apical plasma membranes form a system of leaflets with numerous mitochondria inserted in association with microfilaments. Lateral plasma membranes are linked together by junctional complexes consisting of a zonula adherens and a long convoluted septate junction of the pleated type. In the basal region of the cell, numerous membrane infolds and scattered scalariform junctions with associated mitochondria are present. These cell specializations are typical of arthropod transporting organs, being distinctive features of ion and fluid transporting epithelia. The isolated colon exhibited a transepithelial electrical potential difference (PD) of about 100 mV, lumen side positive with respect to the haemolymph side. The PD was almost abolished by metabolic inhibitors, it was reduced by acetazolamide and SITS, and it was unaffected by ouabain. These effects suggest that HCO3- and Cl- are involved in the genesis of the PD, whereas Na+ is not directly responsible of the PD. Measurements of Na+ and Cl- fluxes across the colon wall confirm that Na+ moves following the PD across the tissue, while Cl- movement occurs against an electrochemical potential difference. The electrical profile of the epithelial cells is of the well type and it suggests that the primary or secondary active step for Cl- transport across the epithelium should be located at the mucosal border of the cell. 相似文献
73.
Samuel H. K. Ho Uttam Das Gupta John S. Rieske 《Journal of bioenergetics and biomembranes》1985,17(5):269-282
Deformamidoazidoantimycin A (DAA), a photoactive derivative of antimycin A containing an azido group substituting for the formamido group attached to the phenyl ring, was synthesized. The ultraviolet spectrum of DAA was almost identical to that of antimycin A, indicating little alteration of the electronic structure of the substituted phenyl ring by the azido substitution. However, the inhibitory effectiveness of DAA toward ubiquinol-cytochromec reductase (Complex III) purified from bovine heart (K
i
=ca. 0.5 µM) was considerably less than that of antimycin (K
i
3 pM), indicating a direct rather than a supporting role of the formamido group in the inhibitory activity of antimycin. Exposure of purified Complex III to [3H]DAA plus ultraviolet light caused a major labeling by tritium of SDS-PAGE band 7 (m=13 kDa by SDS-PAGE) and lesser but significant labeling of bands 3, 6, 8, and 9. Pretreatment of Complex III with antimycin greatly suppressed the labeling of bands 5, 6, and 7 but caused an apparent increased labeling of bands 8 and 9 by [3H]DAA, respectively. The labeling of band 7 by [3H]DAA also was strongly suppressed by reduction of Complex III by either sodium borohybride or ascorbate. Based on magnitude of labeling by [3H]DAA and the degree of suppression of labeling by antimycin, the protein of band 7 qualified as the principal component for specific binding of antimycin with the protein of band 6 (m=16 kDa) showing a lesser but significant amount of specific binding. 相似文献
74.
John T. Christeller Betty E. Terzaghi Diana F. Hill W. A. Laing 《Plant molecular biology》1985,5(4):257-263
Summary The ribulose bisphosphate carboxylase/oxygenase (EC4.1.1.39) (RubisCO) large and small subunit genes from Anacystis nidulans have been cloned as a single fragment into M 13mp10 and pEMBL8 and expressed in Escherichia coli. From M 13mp10 a low yield of enzyme with high specific activity was obtained. The molecular weight of the active enzyme was 260 000 Da and of the inactive enzyme approximately 730 000 Da. The small and large subunits cloned separately did not express activity. The RubisCO gene cloned into pEMBL8 expressed activity up to 22 times that from the M 13 cloned RubisCO DNA. The RubisCO protein produced by the pEMBL cloned gene had a normal MW (550 000). Immunoprecipitation and polyacrylamide gel electrophoresis showed the presence of both large and small subunits. 相似文献
75.
Summary The effect of cholecystokinin (CCK) and internal Ca2+ on outward K+ current in isolated pig pancreatic acinar cells has been investigated using the patch-clamp method for whole-cell current recording under voltage-clamp conditions. CCK (2 × 10–10
M) applied to the bath evoked a marked increase in the outward K+ current associated with depolarizing voltage steps, and this effect was fully reversible and acutely dependent on the presence of external Ca2+. When strongly buffered Ca2+-EGTA solutions were used inside the cells CCK failed to evoke an effect. Increasing the internal Ca2+ concentration ([Ca2+]
i
) from 5 × 10–10
M to 10–7 and 5 × 10–7
M mimicked the effect of CCK. It would appear therefore that CCK controls K+ conductance in the acinar cells via changes in the internal free ionized Ca2+ concentration. 相似文献
76.
Summary Voltage-clamped single nerve fibers of the frogRana esculenta were treated with the carboxyl group activating reagent N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) in the presence of different primary amines and without added amine. Carboxyl groups form stable amide bonds with primary amines in the presence of EEDQ. EEDQ treatment reduced the sodium current considerably and irreversibly, regardless of the presence of a primary amine in the Ringer's solution. The potassium current was also reduced. After modification the reduced sodium currents inactivated slowly and incompletely. The descending branch of the sodium current-voltage relation,I
Na(E), was shifted along the voltage axis in the depolarizing direction. The size of the shift was strongly dependent on the amine present during modification with EEDQ. The voltage-dependence of sodium inactivation,h
x
(E), was shifted to more positive values of membrane potential by EEDQ in the presence of ethylenediamine (11 mV) and glucosamine (3 mV). In contrast, a small shift to more negative potentials occurred in the presence of taurine (–3 mV) or without the addition of an amine (–2 mV). A tenfold increase of the calcium concentration still shifted theI
Na(E) andh
x
(E) curves of the chemically modified fibers. However, these shifts were smaller than those observed on untreated fibers. The currents remaining after the modification were completely blocked by tetrodotoxin; no change of the reversal potential occurred. 相似文献
77.
Abstract: In the present communication we report that Ca2+ -dependent acetylcholine release from K+ -depolarized Torpedo electric organ synaptosomes is inhibited by morphine, and that this effect is blocked by the opiate antagonist naloxone. This finding suggests that the purely cholinergic Torpedo electric organ neurons contain pre-synaptic opiate receptors whose activation inhibits acetylcholine release. The mechanisms underlying this opiate inhibition were investigated by comparing the effects of morphine on acetylcholine release induced by K+ depolarization and by the Ca2+ ionophore A23187 and by examining the effect of morphine on 45 Ca2+ influx into Torpedo nerve terminals. These experiments revealed that morphine inhibits 45 Ca2+ influx into K+ -depolarized Torpedo synaptosomes and that this effect is blocked by naloxone. The effects of morphine on K+ depolarization-mediated 45 Ca2+ influx and on acetylcholine release have similar dose dependencies (half-maximal inhibition at 0.5–1 μ M ), suggesting that opiate inhibition of release is due to blockage of the presynaptic voltage-dependent Ca2+ channel. This conclusion is supported by the finding that morphine does not inhibit acetylcholine release when the Ca2+ channel is bypassed by introducing Ca2+ into the Torpedo nerve terminals via the Ca2+ ionophore. 相似文献
78.
Kodavanti S. Prasada Rao Sreeramulu C. Chetty Durisala Desaiah 《Journal of biochemical and molecular toxicology》1987,2(2):125-140
Tricyclohexylhydroxytin, commonly known as Plictran® inhibited Na+, K+ -ATPase activity of rat brain synaptosomes in a concentration-dependent manner with median inhibitory concentration (IC-50) of 2 μM. Both K+ -stimulated para-nitrophenylphosphatase and [3-H]-ouabain binding to synaptosomes were also inhibited by Plictran with IC-50 values of 11 and 30 μM, respectively. Altered pH and Na+, K+ -ATPase activity curves demonstrated comparable inhibition in buffered neutral and alkaline pH ranges, and no inhibition was observed in acidic pH. The inhibition of Na+, K+ -ATPase was independent of temperature. Kinetic studies of substrate (ATP) activation of Na+, K+ -ATPase indicated uncompetitive inhibition. Results also showed noncompetitive inhibition for p-nitrophenylphosphate and uncompetitive inhibition for K+ activations of p-nitrophenylphosphatase. Preincubation of synaptosomes with dithiothreitol, a sulfhydryl (SH) agent, resulted in the complete protection of Plictran inhibition of Na+, K+ -ATPase, K+ -para-nitrophenylphosphatase, and [3-H]-ouabain binding. The protection was specific and concentration dependent since cysteine and glutathione did not afford protection. These results indicate that Plictran inhibited Na+, K+ -ATPase by interacting with dephosphorylation of the enzyme-phosphoryl complex and exerted a similar effect to that of SH-blocking agents. 相似文献
79.
The Ca2+ indicator 7-chlorotetracycline has been shown to bind to a pore complex on both outer surfaces of all non-meristematic cells in the unistratose thallus of Riella (chlorotetracycline-binding surface region=CSR; Grotha, 1983, Planta 158, 473–481). Prolonged treatment of the thallus with 7-chlorotetracycline, 5-hydroxytetracycline, verapamil and desmethoxyverapamil induces the deposition of callose at the same region. The influence of various treatments on verapamil-induced CSR-callose was measured in situ by microfluorometry of aniline-blue-stained material. Callose deposition is maximal at 10-4M verapamil or 5·10-5M desmethoxyverapamil with 2·10-4M Ca2+ or Mg2+ in the medium. The reaction is completely inhibited at pH 5.5 and is optimal between pH 6.5 and 7.5. The production of CSR-callose is absolutely light-dependent with callose being first visible after 30 min of light. La3+, ethylene glycol-bis(-aminoethylether)-N,N,N,N-tetraacetic acid and amiprophosmethyl, antagonists of Ca2+ functions, and 2-deoxy-D-glucose suppress the verapamil induction of CSR-callose. Furthermore the ionophores A 23187, valinomycin and monensin effectively block the reaction. The deposition of CSR-callose is diminished at increasing external osmolarity and is abolished at osmotic values that stimulate plasmolysis-callose. Wounding causes the formation of wound-callose but inhibits the induction of CSR-callose in cells of the wound edge. Nifedipine increases or prolongs callose synthesis in cell plates. The Ca2+-channel blocker diltiazem is completely ineffective. It is suggested as a working hypothesis that verapamil-induced CSR-callose synthesis is caused by a local change in membrane permeability, possibly as a consequence of the opening of Ca2+ channels being involved in Golgi-vesicle mediated exocytosis (A. Kramer and H. Lehmann, 1986, Ber. Dtsch. Bot. Ges. 99, 111–121).Abbreviations APM
amiprophosmethyl
- APW
artificial pond water
- CSR
chlorotetracycline-binding surface region
- CTC
7-chlorotetracycline
- DDG
2-deoxy-D-glucose
- EGTA
ethylone glycol-bis(2-aminoethylether)-N,N,N,N-tetraacetic acid
- OTC
5-hydroxytetracycline
- Pipes
1,4-piperazinediethane sulfonic acid
Dedicated to Professor Luise Stange on the occasion of her 60th birthday 相似文献
80.
Robert A. MacLeod 《FEMS microbiology letters》1986,39(1-2):109-113
Abstract Many species of bacteria isolated from saline environments require Na+ specifically for membrane transport. Transport occurs by a Na+ symport process energized by an electrochemical gradient of Na+ ions. The gradient at neutral pH appears to be produced by a primary electrogenic extrusion of protons coupled to a secondary, outwardly directed Na+ pump, a Na+ /proton antiporter. At alkaline pH Vibrio alginolyticus may also produce the gradient by an energy-dependent primary extrusion of Na+ ions. Alteromonas haloplanktis and Vibrio costicola require salts in the medium to retain intracellular solutes. For A. haloplanktis the effects of the salts are primarily osmotic. For V. costicola , only NaCl is effective in retaining solutes and Na+ is required by this organism to maintain the membrane potential. In Escherichia coli a single substitution in the nucleotide sequence of the gene coding for the melibiose transport protein changed the cation specificity of the transport system. The possible ecological significance of this finding has been considered. 相似文献