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41.
Mark Tester 《The Journal of membrane biology》1988,105(1):77-85
Summary The outer membranes of plant cells contain channels which are highly selective for K+. In the giant-celled green algaChara corallina, K+ currents in the plasmalemma were measured during the action potential and when the cell was depolarized to the K+ equilibrium potential in high external K+ concentrations. Currents in both conditions were reduced by externally added tetraethylammonium (TEA+), Ba2+, Na+ and Cs+. In contrast to inhibition by TEA+, the latter three ions inhibited inward K+ current in a voltage-dependent manner, and reduced inward current more than outward. Ba2+ and Na+ also appeared to inhibit outward current in a strongly voltage-dependent manner. The blockade by Cs+ is studied in more detail in the following paper. TEA+ inhibited both inward and outward currents in a largely voltage-independent manner, with an apparentK
D
of about 0.7 to 1.1mm, increasing with increasing external K+. All inhibitors reduced current towards a similar linear leak, suggesting an insensitivity of the background leak inChara to these various K+ channel inhibitors. The selectivity of the channel to various monovalent cations varied depending on the method of measurement, suggesting that ion movement through the K+-selective channel may not be independent. 相似文献
42.
43.
Roberta Colombo Raffaella Cerana Piera Lado Antonio Peres 《The Journal of membrane biology》1988,103(3):227-236
Summary The patch-clamp technique in whole-cell configuration was used to study the electrical properties of the tonoplast in isolated vacuoles fromAcer pseudoplatanus cultured cells. In symmetrical KCl or K2 malate solutions, voltage- and time-dependent inward currents were elicited by hyperpolarizing the tonoplast (inside negative), while in the positive range of potential the conductance was very small. The specific conductance of the tonoplast at –100 mV, in 100mm symmetrical KCl was about 160 S/cm2. The reversal potentials (E
rev) of the current, measured in symmetrical or asymmetrical ion concentrations (cation, anion or both) were very close to the values of the K+ equilibrium potential. Experiments performed in symmetrical or asymmetrical NaCl indicate that Na+ too can flow through the channels. NeitherE
rev nor amplitude and kinetics of the current changed by replacing NaCl with KCl in the external solution. These results indicate the presence of hyperpolarization-activated channels in tonoplasts, which are permeable to K+ as well as to Na+. Anions such as Cl– or malate seem to contribute little to the channel current. 相似文献
44.
Summary Quantitative electron microprobe analysis was employed to compare the effects of aldosterone and ADH on the intracellular electrolyte concentrations in the toad urinary bladder epithelium. The measurements were performed on thin freeze-dried cryosections utilizing energy dispersive x-ray microanalysis. After aldosterone, a statistically significant increase in the intracellular Na concentration was detectable in 8 out of 9 experiments. The mean Na concentration of granular cells increased from 8.9±1.3 to 13.2±2.2 mmol/kg wet wt. A significantly larger Na increase was observed after an equivalent stimulation of transepithelial Na transport by ADH. On average, the Na concentration in granular cells increased from 12.0±2.3 to 31.4±9.3 mmol/kg wet wt (5 experiments). We conclude from these results that aldosterone, in addition to its stimulatory effect on the apical Na influx, also exerts a stimulatory effect on the Na pump. Based on a significant reduction in the Cl concentration of granular cells, we discuss the possibility that the stimulation of the pump is mediated by an aldosterone-induced alkalinization.Similar though less pronounced concentration changes were observed in basal cells, suggesting that this cell type also participates in transepithelial Na transport. Measurements in mitochondria-rich cells provided no consistent results. 相似文献
45.
符云峰 《中国生物化学与分子生物学报》1988,4(1):84-87
本研究确定了在0℃条件下,(Na~++K~+)-ATP酶纯化制备物与5mmol/L Na~+或Mg~(2+)在5mmol/L咪唑(pH7.4)环境中预保温30分钟,然后进行磷酸化,可以获得最高磷酸化水平,Na~+或Mg~(2+)的K_(0.5)值分别为0.29mmol/L或0.35mmol/L;以ADP代替Na~+和Mg~(2+)与酶预保温,对E_2向E_1转变无任何影响,而与Na~+、Mg~(2+)一起存在时则能加强Na~+及Mg~2的预保温效果。 相似文献
46.
47.
haemolysin of Escherichia coli: Comparison of pore-forming properties between chromosome and plasmid-encoded haemolysins 总被引:3,自引:0,他引:3
Roland Benz reas Döbereiner Albrecht Ludwig Werner Goebel 《FEMS microbiology letters》1992,105(1-3):55-62
Abstract Lipid bilayer experiments were performed with chromosome-encoded haemolysin of Escherichia coli . The addition of the toxin to the aqueous phase bathing lipid bilayer membranes of asolectin resulted in the formation of transient ion-permeable channels with two states at small transmembrane voltages. One is prestate (single-channel conductance 40 pS in 0.15 M KCl) of the open state, which had a single-channel conductance of 420 pS in 0.15 M KCl and a mean lifetime of 30 s. Membranes formed of pure lipids were rather inactive targets for this haemolysin. Experiments with different salts suggested that the haemolysin channel was highly cation-selective at neutral pH. The mobility sequence of the cations in the channel was similar if not identical to their mobility sequence in the aqueous phase. The single-channel data were consistent with a wide, water-filled channel with an estimated minimal diameter of about 1 nm. The pore-forming properties of chromosome-encoded haemolysin were compared with those of plasmid-encoded haemolysin. Both toxins share common features, oligomerize probably to form pores in lipid bilayer membranes. Both types of haemolysin channels have similar properties but different lifetimes. 相似文献
48.
1. Two mutants of the sodium channel II have been expressed inXenopus oocytes and have been investigated using the patch-clamp technique. In mutant E387Q the glutamic acid at position 387 has been replaced by glutamine, and in mutant D384N the aspartic acid at position 384 has been replaced by asparagine.2. Mutant E387Q, previously shown to be resistant to block by tetrodotoxin (Noda et al. 1989), has a single-channel conductance of 4 pS, that can be easily measured only using noise analysis. At variance with the wild-type, the openchannel current-voltage relationship of mutant E387Q is linear over a wide voltage range even under asymmetrical ionic conditions.3. Mutant D384N has a very low permeability for any of the following ions: Cl–, Na+, K+, Li+, Rb+, Ca2+, Mg2+, NH4
+ , TMA+, TEA+. However, asymmetric charge movements similar to the gating currents of the Na+-selective wild-type are still observed.4. These results suggest that residues E387 and D384 interact directly with the pathway of the ions permeating the open channel.Abbreviations TTX
tetrodotoxin; Na+, sodium; K+, potassium;
- NFR
normal frog Ringer
- HEPES
N-2-hydroxylethyl piperazine-N-2-ethanesulfonic acid
- EGTA
ethyleneglycol-bis(-amino-ethyl ether) N,N,N',N'-tetra acetic acid
- TEA
tetraethylammonium
- TMA
tetramethylammonium;I
g
, gating current; , single-channel conductance 相似文献
49.
Michiaki Morohashi Keiko Tsuchiya Takashi Mita Masaru Kawamura 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(1):69-72
Summary An inhibitory activity to (Na,K)ATPase was found in cell extracts of the brine shrimp, Artemia salina, irrespective of its developmental stages. Organic solvent extraction together with gas chromatographic analysis reveals that the inhibitory activity is due to long-chain, non-esterified fatty acids and their derivatives. Unsaturated fatty acids, especially with cis-configuration, are more effective in inhibition than saturated ones.Abbreviations ATPase
adenosine triphosphatase
- EDTA
ethylenediamine-tetraacetate
- TLC
thin-layer chromatography 相似文献
50.
The inside-out mode of the patch-clamp technique was used to study adenosine-5-triphosphate (ATP)-sensitive K+ channels in mammalian skeletal muscle. Vanadate, applied to the cytoplasmic face of excised patches, was a potent activator of ATP-sensitive K+ channels. Divalent cations (Mg2+, Ca2+) were a prerequisite for the activating process. The maximal effect was achieved using 1 mM vanadate dissolved in Ringer, increasing the open-state probability about ninefold. The active 5 + redox form of vanadate which stimulates ATP-sensitive K+ channels is likely to be decavanadate V10O
inf28
sup6–
. ATP concentration-response curves have Hill coefficients near three in internal Na+-rich Ringer and between one and two in internal KCl solutions. Half maximal channel blockage was observed at ATP concentrations of 4 and 8 M in Ringer and KCl solutions, respectively. Internal vanadate shifted the curves towards higher ATP concentrations without affecting their slopes. Thus 50% channel blockage occurred at 65 M ATP in internal Ringer containing 0.5 mM vanadate. The results indicate that the affinity and stoichiometry of ATP binding to ATP-sensitive K+ channels are strongly modulated by internal cations and that the ATP sensitivity is weakened by vanadate.
Offprint requests to: B. Neumcke 相似文献