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1.
Stanley J. Roux 《植物生理与分子生物学学报》2014,(6):937-938
A START POINT FOR EXTRACELLULAR NUCLEOTIDE SIGNALING
The recent discovery of a plant receptor for extracellu- lar nucleotides, reported by Choi et al. (2014), is a major breakthrough that had been anticipated for over a dec- ade. Plants release ATP into their extracellular matrix (ECM) during growth and when they are induced by vari- ous biotic and abiotic stimuli (Clark and Roux, 2011). That these extracellular nucleotides would activate receptors in plants was predicted by two sets of discoveries: that low- and sub-micromolar ATP could induce increases in [Ca2+]cyt, NO, and superoxide signaling intermediates that led to downstream growth, stomatal, and defense responses, and that these changes could be blocked by antagonists that blocked extracellular nucleotide receptors in animals (Demidchik et al., 2003; Song et al., 2006; Clark et al., 2011; Demidchik et al., 2009, 2011). Although mammalian biolo- gists had discovered two classes of receptors for extracel- lular nucleotides (P2X and P2Y) decades ago (Burnstock, 2007), there were no plant proteins obviously similar to these in any sequence data available. Clearly, if there were plant purinoceptors, they would be different from the mammalian receptors, and they could not be discovered by motif searches. 相似文献
The recent discovery of a plant receptor for extracellu- lar nucleotides, reported by Choi et al. (2014), is a major breakthrough that had been anticipated for over a dec- ade. Plants release ATP into their extracellular matrix (ECM) during growth and when they are induced by vari- ous biotic and abiotic stimuli (Clark and Roux, 2011). That these extracellular nucleotides would activate receptors in plants was predicted by two sets of discoveries: that low- and sub-micromolar ATP could induce increases in [Ca2+]cyt, NO, and superoxide signaling intermediates that led to downstream growth, stomatal, and defense responses, and that these changes could be blocked by antagonists that blocked extracellular nucleotide receptors in animals (Demidchik et al., 2003; Song et al., 2006; Clark et al., 2011; Demidchik et al., 2009, 2011). Although mammalian biolo- gists had discovered two classes of receptors for extracel- lular nucleotides (P2X and P2Y) decades ago (Burnstock, 2007), there were no plant proteins obviously similar to these in any sequence data available. Clearly, if there were plant purinoceptors, they would be different from the mammalian receptors, and they could not be discovered by motif searches. 相似文献
2.
Two mechanisms of passive Ca2+ transport, Na+-Ca2+ exchange and Ca2+-Ca2+ exchange, were studied using highly-purified dog heart sarcolemmal vesicles. About 80% of the Ca2+ accumulated by Na+-Ca2+ exchange or Ca2+-Ca2+ exchange could be released as free Ca2+, while up to 20% was probably bound. Na+-Ca2+ exchange was simultaneous, coupled countertransport of Na+ and Ca2+. The movement of anions during Na+-Ca2+ exchange did not limit the initial rate of Na+-Ca2+ exchange. Na+-Ca2+ exchange was electrogenic, with a reversal potential of about -105 mV. The apparent flux ratio of Na+-Ca2+ exchange was 4 Na+:1 Ca2+. Coupled cation countertransport by the Na+-Ca2+ exchange mechanism required a monovalent cation gradient with the following sequence of ion activation: Na+ much greater than Li+ greater than Cs+ greater than K+ greater than Rb+. In contrast to Na+-Ca2+ exchange, Ca2+-Ca2+ exchange did not require a monovalent cation gradient, but required the presence of Ca2+ plus a monovalent cation on both sides of the vesicle membrane. The sequence of ion activation of Ca2+-Ca2+ exchange was: K+ much greater than Rb+ greater than Na+ greater than Li+ greater than Cs+. Na+ inhibited Ca2+-Ca2+ exchange when Ca2+-Ca2+ exchange was supported by another monovalent cation. Both Na+-Ca2+ exchange and Ca2+-Ca2+ exchange were inhibited, but with different sensitivities, by external MgCl2, quinidine, or verapamil. 相似文献
3.
Natalia Von Ellenrieder 《Systematic Entomology》2002,27(4):437-467
Abstract A cladistic analysis of the world Aeshnidae is presented, based on fifty-eight characters of adult and larval anatomy. The ingroup taxa include all the extant genera of Aeshnidae, and the austropetaliid genera Phyllopetalia and Hypopetalia were chosen as the outgroup. The strict consensus tree obtained after successive weighting shows that the subgroups defined traditionally for Aeshnidae are paraphyletic or polyphyletic. The previous reclassification derived from analyses based on wing venation is supported in terms of the monophyly of Aeshnidae, Gomphaeschninae and its sister group comprising the remaining Aeshnidae. Gomphaeschninae is confirmed as sister group of the remaining Aeshnidae (= Aeshnodea Bechly). The sister-group relationships between Gomphaeschna + Sarasaeschna and Linaeschna + Oligoaeschna are corroborated. Within Aeshnodea, three monophyletic groups emerged: Boyeria + ( Petaliaeschna + ( Limnetron + Gynacanthaeschna + Periaeschna )) + (( Cephalaeschna + Caliaeschna ) + ( Allopetalia ( Notoaeschna + Spinaeschna ))); Dendroaeschna + ( Epiaeschna + ( Aeschnophlebia + ( Nasiaeschna + ( Tetracanthagyna + Brachytron )))); and Polycanthagyna + ( Basiaeschna + ( Amphiaeschna + ( Indaeschna + ( Oplonaeschna + ( Racenaeschna + Plattycantha + Agyrtacantha + Triacanthagyna + ( Subaeschna + Austrogynacantha + Gynacantha ) + ( Heliaeschna + ( Neuraeschna + Staurophlebia ))) + (( Castoraeschna + Coryphaeschna + Remartinia ) + ( Oreaeschna + ( Aeshna + ( Anaciaeschna + ((' A .' isosceles + Andaeshna ) + ( Anax + Hemianax )))))). Additional informative characters are required to test the relationships suggested here between the main groups of Aeshnodea and some enigmatic basal taxa ( Antipodophlebia , Austroaeschna , Acanthaeschna , Telephlebia , Austrophlebia and Planaeschna ). 相似文献
4.
Dunham PB Kelley SJ Logue PJ Mutolo MJ Milanick MA 《American journal of physiology. Cell physiology》2005,289(2):C277-C282
Amiloride-inhibitable Li+ influx in dog red blood cells is mediated by the Na+/H+ exchanger, NHE. However, there are substantial differences between the properties of Li+ transport and Na+ transport through the NHE. Li+ influx is activated by cell shrinkage, and Na+ influx is not, as we reported previously (Dunham PB, Kelley SJ, and Logue PJ. Am J Physiol Cell Physiol 287: C336-C344, 2004). Li+ influx is a sigmoidal function of its concentration, and Na+ activation is linear at low Na+ concentrations. Li+ does not inhibit its own influx; in contrast, Na+ inhibits Na+ influx. Li+ prevents this inhibition by Na+. Na+ is a mixed or noncompetitive inhibitor of Li+ influx, implying that both a Na+ and a Li+ can be bound at the same time. In contrast, Li+ is a competitive inhibitor of Na+ influx, suggesting Li+ binding at one class of sites on the transporter. Because the properties of Li+ transport and Na+ transport are different, a simple explanation is that Na+ and Li+ are transported by separate sites. The similarities of the properties of Li+ transport and the inhibition of Na+ transport by Na+ suggest that Li+ is transported by the Na+-inhibitory sites. 相似文献
5.
Showdomycin inhibited pig brain (Na+ + K+)-ATPase with pseudo first-order kinetics. The rate of inhibition by showdomycin was examined in the presence of 16 combinations of four ligands, i.e., Na+, K+, Mg2+ and ATP, and was found to depend on the ligands added. Combinations of ligands were divided into five groups in terms of the magnitude of the rate constant; in the order of decreasing rate constants these were: (1) Na+ + Mg2+ + ATP, (2) Mg2+, Mg2+ + K+, K+ and none, (3) Na+ + Mg2+, Na+, K+ + Na+ and Na+ + K+ + Mg2+, (4) Mg2+ + K+ + ATP, K+ + ATP and Mg2+ + ATP, (5) K+ + Na + + ATP, Na+ + ATP, Na+ + K+ + Mg2+ + ATP and ATP. The highest rate was obtained in the presence of Na+, Mg2+ and ATP. The apparent concentrations of Na+, Mg2+ and ATP for half-maximum stimulation of inhibition (KS0.5) were 3 mM, 0.13 mM and 4 MicroM, respectively. The rate was unchanged upon further increase in Na+ concentration from 140 to 1000 mM. The rates of inhibition could be explained on the basis of the enzyme forms present, including E1, E2, ES, E1-P and E2-P, i. e., E2 has higher reactivity with showdomycin than E1, while E2-P has almost the same reactivity as E1-P. We conclude that the reaction of (Na+ + K+)- ATPase proceeds via at least four kinds of enzyme form (E1, E2, E1 . nucleotide and EP), which all have different conformations. 相似文献
6.
Qing Wang Bi Ma Xiwu Qi Qing Guo Xuwei Wang Qiwei Zeng Ningjia He 《植物学报(英文版)》2014,56(7):663-672
Jasmonate (JA) is an important phytohormone regulating growth, development, and environmental response in plants, particularly defense response against herbivorous insects. Recently, completion of the draft genome of the mulberry (Morus notabilis) in conjunction with genome sequencing of silkworm (Bombyx mori) provides an opportuni-ty to study this unique plant-herbivore interaction. Here, we identified genes involved in JA biosynthetic and signaling pathways in the genome of mulberry for the first time, with the majority of samples showing a tissue-biased expression pattern. The analysis of the representative genes 12-oxophy-todienoic acid reductase (OPRs) and jasmonate ZIM-domain (JAZs) was performed and the results indicated that the mulberry genome contains a relatively smal number of JA biosynthetic and signaling pathway genes. A gene encoding an important repressor, MnNINJA, was identified as an alternative splicing variant lacking an ethylene-responsive element binding factor-associated amphiphilic repression motif. Having this fundamental information wil facilitate future functional study of JA-related genes pertaining to mulberry-silkworm interactions. 相似文献
7.
Rat liver 3-hydroxy-3-methylglutaryl-CoA reductase. Catalysis of the reverse reaction and two half-reactions 总被引:1,自引:0,他引:1
D G Sherban P J Kennelly K G Brandt V W Rodwell 《The Journal of biological chemistry》1985,260(23):12579-12585
Rat liver 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase catalyzes, in addition to its normal biosynthetic or forward reaction (HMG-CoA + 2 NADPH + 2H+----mevalonate + 2 NAD+ + CoASH), the reverse reaction (mevalonate + CoASH + 2 NADP+----HMG-CoA + 2 NADPH + 2H+) and two "half-reactions" that involve the presumed intermediate mevaldate (mevaldate + CoASH + NADP+----HMG-CoA + NADPH + H+ and mevaldate + NADPH + H+----mevalonate + NADP+). These reactions were studied using both enzyme solubilized by the traditional freeze-thaw method and enzyme solubilized with a nonionic detergent in the presence of inhibitors of proteolysis. All four reactions were inhibited by mevinolin, a known inhibitor of the forward (biosynthetic) reaction catalyzed by HMG-CoA reductase. When the enzyme was inactivated by ATP and a cytosolic, ADP-dependent HMG-CoA reductase kinase, the rates of both the forward reaction and the half-reactions decreased to comparable extents. Although coenzyme A is not a stoichiometric participant in the second half-reaction (mevaldate + NADPH + H+----mevalonate + NADP+), it was required as an activator of this reaction. This observation implies that coenzyme A may remain bound to the enzyme throughout the normal catalytic cycle of HMG-CoA reductase. 相似文献
8.
The effect of Mg2+ on hepatic microsomal Ca2+ and Sr2+ transport 总被引:2,自引:0,他引:2
The ATP-dependent uptake of Ca2+ by rat liver microsomal fraction is dependent upon Mg2+. Studies of the Mg2+ requirement of the underlying microsomal Ca2+-ATPase have been hampered by the presence of a large basal Mg2+-ATPase activity. We have examined the effect of various Mg2+ concentrations on Mg2+-ATPase activity, Ca2+ uptake, Ca2+-ATPase activity and microsomal phosphoprotein formation. Both Mg2+-ATPase activity and Ca2+ uptake were markedly stimulated by increasing Mg2+ concentration. However, the Ca2+-ATPase activity, measured concomitantly with Ca2+ uptake, was apparently unaffected by changes in the Mg2+ concentration. In order to examine the apparent paradox of Mg2+ stimulation of Ca2+ uptake but not of Ca2+-ATPase activity, we examined the formation of the Ca2+-ATPase phosphoenzyme intermediate and formation of a Mg2+-dependent phosphoprotein, which we have proposed to be an attribute of the Mg2+-ATPase activity. We found that Ca2+ apparently inhibited formation of the Mg2+-dependent phosphoprotein both in the absence and presence of exogenous Mg2+. This suggests that Ca2+ may inhibit (at least partially) the Mg2+-ATPase activity. However, inclusion of the Ca2+ inhibition of Mg2+-ATPase activity in the calculation of Ca2+-ATPase activity reveals that this effect is insufficient to totally account for the stimulation of Ca2+ uptake by Mg2+. This suggests that Mg2+, in addition to stimulation of Ca2+-ATPase activity, may have a direct stimulatory effect on Ca2+ uptake in an as yet undefined fashion. In an effort to further examine the effect of Mg2+ on the microsomal Ca2+ transport system of rat liver, the interaction of this system with Sr2+ was examined. Sr2+ was sequestered into an A23187-releasable space in an ATP-dependent manner by rat liver microsomal fraction. The uptake of Sr2+ was similar to that of Ca2+ in terms of both rate and extent. A Sr2+-dependent ATPase activity was associated with the Sr2+ uptake. Sr2+ promoted formation of a phosphoprotein which was hydroxylamine-labile and base-labile. This phosphoprotein was indistinguishable from the Ca2+-dependent ATPase phosphoenzyme intermediate. Sr2+ uptake was markedly stimulated by exogenous Mg2+, but the Sr2+-dependent ATPase activity was unaffected by increasing Mg2+ concentrations. Sr2+ uptake and Sr2+-dependent ATPase activity were concomitantly inhibited by sodium vanadate. In contrast to Ca2+, Sr2+ had no effect on Mg2+-dependent phosphoprotein formation. Taken together, these data indicate that Mg2+ stimulated Ca2+ and Sr2+ transport by increasing the Ca2+ (Sr2+)/ATP ratio.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
9.
The involvement of membrane (Na+ + K+)-ATPase (Mg2+-dependent, (Na+ + K+)-activated ATP phosphohydrolase, E.C. 3.6.1.3) in the oxygen consumption of rat brain cortical slices was studied in order to determine whether (Na+ + K+)-ATPase activity in intact cells can be estimated from oxygen consumption. The stimulation of brain slice respiration with K+ required the simultaneous presence of Na+. Ouabain, a specific inhibitor of (Na+ + K+)-ATPase, significantly inhibited the (Na+ + K+)-stimulation of respiration. These observations suggest that the (Na+ + K+)-stimulation of brain slice respiration is related to ADP production as a result of (Na+ + K+)-ATPase activity. However, ouabain also inhibited non-K+ -stimulated respiration. Additionally, ouabain markedly reduced the stimulation of respiration by 2,4-dinitrophenol in a high (Na+ + K+)-medium. Thus, ouabain depresses brain slice respiration by reducing the availability of ADP through (Na+ + K+)-ATPase inhibition and acts additionally by increasing the intracellular Na+ concentration. These studies indicate that the use of ouabain results in an over-estimation of the respiration related to (Na+ + K+)-ATPase activity. This fraction of the respiration can be estimated more precisely from the difference between slice respiration in high Na+ and K+ media and that in choline, K+ media. Studies were performed with two (Na+ + K+)-ATPase inhibitors to determine whether administration of these agents to intact rats would produce changes in brain respiration and (Na+ + K+)-ATPase activity. The intraperitoneal injection of digitoxin in rats caused an inhibition of brain (Na+ + K+)-ATPase and related respiration, but chlorpromazine failed to alter either (Na+ + K+)-ATPase activity or related respiration. 相似文献
10.
Paydar MJ Pousti A Farsam H Amanlou M Mehr SE Dehpour AR 《Canadian journal of physiology and pharmacology》2005,83(11):967-975
The purpose of this study was to determine the effects of 2 Ca2+ channel blockers, verapamil and diltiazem, on calcium loading (active Ca2+ uptake) and the following Ca2+ release induced by silver ion (Ag+) and Ca2+ from the membrane of heavy sarcoplasmic reticulum (SR) of chicken skeletal muscle. A fluorescent probe technique was employed to determine the calcium movement through the SR. Pretreatment of the medium with diltiazem and verapamil resulted in a significant decrease in the active Ca2+ uptake, with IC50 of about 290 micromol/L for verapamil and 260 micromol/L for diltiazem. Inhibition of Ca2+ uptake was not due to the development of a substantial drug-dependent leak of Ca2+ from the SR. It might, in part, have been mediated by a direct inhibitory effect of these drugs on the Ca2+ ATPase activity of the SR Ca2+ pump. We confirmed that Ca2+ channel blockers, administered after SR Ca2+ loading and before induction of Ca2+ release, caused a dose-dependent inhibition of both Ca2+- and Ag+-induced Ca2+ release rate. Moreover, if Ca2+ channel blockers were administered prior to SR Ca2+ loading, in spite of Ca2+ uptake inhibition the same reduction in Ca2+- and Ag+-induced Ca2+ release rate was seen. We showed that the inhibition of Ag+-induced Ca2+ release by L-channel blockers is more sensitive than Ca2+-induced Ca2+ release inhibition, so the IC50 for Ag+- and Ca2+-induced Ca2+ release was about 100 and 310 micromol/L for verapamil and 79 and 330 micromol/L for diltiazem, respectively. Our results support the evidence that Ca2+ channel blockers affect muscle microsome of chicken skeletal muscle by 2 independent mechanisms: first, reduction of Ca2+ uptake rate and Ca2+-ATPase activity inhibition, and second, inhibition of both Ag+- and Ca2+-induced Ca2+ release by Ca2+ release channels. These findings confirm the direct effect of Ca2+ channel blockers on calcium release channels. Our results suggest that even if the SR is incompletely preloaded with Ca2+ because of inhibition of Ca2+ uptake by verapamil and diltiazem, no impairment in Ca2+ release occurs. 相似文献
11.
We tested the hypothesis that Na+ uptake and H+ release at fertilization of Urechis eggs might occur via a Na+:H+ exchange. Previous studies have shown that (1) Na+ uptake is proportional to the number of entering sperm in seawater with or without lowered Na+ and (2) H+ release is proportional to external pH. Therefore, to determine if Na+ uptake and H+ release are always proportional, we determined the effect of polyspermy on H+ release in natural and low Na+ seawater and the effect of external pH on Na+ uptake and release. Na+ uptake and H+ release do not covary in a manner consistent with a Na+:H+ exchange. H+ release under most conditions was manner consistent with a Na+:H+ exchange. H+ release under most conditions was independent of the number of sperm/egg and in low Na+ seawater was at most 53 +/- 16% of that in natural seawater. In contrast, Na+ uptake in low Na+ seawater can be more than in natural seawater (Jaffe et al., J. Gen. Physiol. 73, 469-492, 1979). In natural seawater Na+ uptake exceeded H+ release; at pH 7 Na+ uptake was 2 pmol/egg, but there was no H+ release. Since Na+ release did not increase at fertilization at pH 7, neither Na+:Na+ nor Na+:H+ exchange could account for the Na+ uptake. An alternate hypothesis is suggested: Na+ uptake is primarily via the channels responsible for the fertilization potential, while H+ release is by another route that is affected by the membrane potential during the fertilization potential. 相似文献
12.
Periodic cyclic-AMP pulses control the cell aggregation and differentiation of Dictyostelium discoideum. Another component required for the aggregation and differentiation of these cells appears to be extracellular Ca+ +. Oscillations in extracellular Ca+ + concentration were investigated in suspensions of differentiating cells. We observed spike-shaped and sinusoidal Ca+ + oscillations. In the course of differentiation, spike-shaped Ca+ + oscillations preceded sinusoidal oscillations, and no phase change occurred at the transition from spike-shaped to sinusoidal Ca+ + oscillations. Spike-shaped and sinusoidal Ca+ + oscillations were related to oscillations in (1) the cyclic-AMP and cyclic-GMP content of cells, (2) the light-scattering properties of cells, and (3) the extracellular pH. Spikeshaped Ca+ + oscillations were observed together with cyclic-AMP oscillations. The minima of the extracellular Ca+ + concentration trailed the maxima of the cyclic-AMP concentration by about 30 s. Sinusoidal Ca+ + oscillations were not accompanied by measurable cyclic-AMP oscillations. The amplitudes of the sinusoidal Ca+ + oscillations were smaller than those of the spike-shaped Ca+ + oscillations. A Ca+ + oscillation of small amplitude (instead of a spike-shaped oscillation) was observed when one cyclic-AMP spike was skipped. Our results provide evidence for the existence of a sinusoidal cyclic-AMP-independent Ca+ + oscillation of small amplitude, and they also suggest that spike-shaped Ca+ + oscillations may be superimposed on such small-amplitude oscillations. When D. discoideum cells produce cyclic-AMP spikes, the uptake of additional Ca+ + is induced, resulting in Ca+ + oscillations of a large amplitude. 相似文献
13.
S M Periyasamy S S Kakar K D Garlid A Askari 《The Journal of biological chemistry》1990,265(11):6035-6041
In bovine cardiac sarcolemmal vesicles, an outward H+ gradient stimulated the initial rate of amiloride-sensitive uptake of 22Na+, 42K+, or 86Rb+. Release of H+ from the vesicles was stimulated by extravesicular Na+, K+, Rb+, or Li+ but not by choline or N-methylglucamine. Uptakes of Na+ and Rb+ were half-saturated at 3 mM Na+ and 3 mM Rb+, but the maximal velocity of Na+ uptake was 1.5 times that of Rb+ uptake. Na+ uptake was inhibited by extravesicular K+, Rb+, or Li+, and Rb+ uptake was inhibited by extravesicular Na+ or Li+. Amiloride-sensitive uptake of Na+ or Rb+ increased with increase in extravesicular pH and decrease in intravesicular pH. In the absence of pH gradient, there were stimulations of Na+ uptake by intravesicular Na+ and K+ and of Rb+ uptake by intravesicular Rb+ and Na+. Similarly, there were trans stimulations of Na+ and Rb+ efflux by extravesicular alkali cations. The data suggest the existence of a nonselective antiporter catalyzing either alkali cation/H+ exchange or alkali cation/alkali cation exchange. Since increasing Na+ caused complete inhibition of Rb+/H+ exchange, but saturating K+ caused partial inhibitions of Na+/H+ exchange and Na+/Na+ exchange, the presence of a Na(+)-selective antiporter is also indicated. Although both antiporters may be involved in pH homeostasis, a role of the nonselective antiporter may be in the control of Na+/K+ exchange across the cardiac sarcolemma. 相似文献
14.
Oki Iwashiro Tamura Sachiko Nishino Machiko Takai Masayuki Kuznedelov Konstantin D. Timoshkin Oleg A. Kawakatsu Masaharu 《Hydrobiologia》1998,383(1-3):315-320
Two lake-dwelling species of paludicolen triclads from Lake Biwa-ko (Honshû, Japan) were studied taxonomically and karyologically. (1) Phagocata kawakatsui Okugawa, 1956, is an epigean species usually inhabiting shallow springs and spring-fed streams in Central Japan. In Lake Biwa-ko, animals were obtained from several bottom stations of the littoral area in the southern part of the northern basin (3–70 m in depth). Chromosome numbers and karyotype: 2x=24 (2m+2sm+2sm+2m+2sm+2m+2sm+2m+2m+2sm+2m+2m). The first pair of metacentric chromosomes is very large in size. (2) Bdellocephala annandalei Ijima et Kaburaki, 1916, an endemic species, is distributed widely in the deep areas of the northern basin (30 to over 100 m in depth). Chromosome numbers and karyotype: 2x=28 (2m+2sm+2sm+2sm+2sm+2m+2m+2m+2m+2m+2m+2m+2m+2m) with the first pair of metacentric chromosomes very long. 相似文献
15.
Crambert G Horisberger JD Modyanov NN Geering K 《American journal of physiology. Cell physiology》2002,283(1):C305-C314
To investigate whether nongastric H+-K+-ATPases transport Na+ in exchange for K+ and whether different beta-isoforms influence their transport properties, we compared the functional properties of the catalytic subunit of human nongastric H+-K+-ATPase, ATP1al1 (AL1), and of the Na+-K+-ATPase alpha1-subunit (alpha1) expressed in Xenopus oocytes, with different beta-subunits. Our results show that betaHK and beta1-NK can produce functional AL1/beta complexes at the oocyte cell surface that, in contrast to alpha1/beta1 NK and alpha1/betaHK complexes, exhibit a similar apparent K+ affinity. Similar to Na+-K+-ATPase, AL1/beta complexes are able to decrease intracellular Na+ concentrations in Na+-loaded oocytes, and their K+ transport depends on intra- and extracellular Na+ concentrations. Finally, controlled trypsinolysis reveals that beta-isoforms influence the protease sensitivity of AL1 and alpha1 and that AL1/beta complexes, similar to the Na+-K+-ATPase, can undergo distinct K+-Na+- and ouabain-dependent conformational changes. These results provide new evidence that the human nongastric H+-K+-ATPase interacts with and transports Na+ in exchange for K+ and that beta-isoforms have a distinct effect on the overall structural integrity of AL1 but influence its transport properties less than those of the Na+-K+-ATPase alpha-subunit. 相似文献
16.
毛百合根尖染色体Giemsa C-带分析 总被引:1,自引:0,他引:1
本研究利用Giemsa C-带方法对毛百合(Lilium dahuricum Ker-Gawl)根尖染色体进行了分析。研究结果表明毛百合试管苗的染色体倍性变异丰富,染色体倍性变异包括二倍体(2n=2×=24)、三倍体(2n=3×=36)、四倍体(2n=4×=48)到六倍体(2n=6×=72)。对二倍体毛百合的C-带结果进行分析,其带型公式为:2n=2×=24=2CI++2CI+T+T++6I+2I++2I++2I+T++2I+T++2I+T++2T++2T+。每条染色体上都显示出显著的特征带,而且带纹的深浅差异明显。强带主要集中在长短臂上。因此,GiemsaC-带方法可以将毛百合(L.dahuricum)的每条染色体区分开。 相似文献
17.
The role of intracellular Ca2+ as essential activator of the Na+-Ca2+ exchange carrier was explored in membrane vesicles containing 67% right-side-out and 10% inside-out vesicles, isolated from squid optic nerves. Vesicles containing 100 microM free calcium exhibited a 2-fold increase in the initial rate of Na+i-dependent Ca2+ uptake as compared with vesicles where intravesicular calcium was chelated by 2 mM EGTA or 10 mM HEDTA. The activatory effect exerted by intravesicular Ca2+ on the reverse mode of Na+-Ca2+ exchange (i.e. Na+i-Ca2+o exchange) is saturated at about 100 microM Ca2+i and displays an apparent K 1/2 of 12 microM. Intravesicular Ca2+ produced activation of Na+i-Ca2+i exchange activity rather than an increase in Ca2+ uptake due to Ca2+-Ca2+ exchange. The presence of Ca2+i was essential for the Na+i-dependent Na+ influx, a partial reaction of the Na+-Ca2+ exchanger. In fact, the Na+ influx levels in vesicles loaded with 2 mM EGTA were close to those expected from diffusional leak while in vesicles containing Ca2+i an additional Na+-Na+ exchange was measured. The results suggest that in nerve membrane vesicles Ca2+ at the inner aspect of the membrane acts as an activator of the Na+-Ca2+ exchange system. 相似文献
18.
In experiments performed at 37 degrees C, Ca2+ reversibly inhibits the Na+-and (Na+ + K+)-ATPase activities and the K+-dependent phosphatase activity of (Na+ + K+)-ATPase. With 3 mM ATP, the Na+-ATPase was less sensitive to CaCl2 than the (Na+ + K+)-ATPase activity. With 0.02 mM ATP, the Na+-ATPase and the (Na+ + K+)-ATPase activities were similarly inhibited by CaCl2. The K0.5 for Ca2+ as (Na+ + K+)-ATPase inhibitor depended on the total MgCl2 and ATP concentrations. This Ca2+ inhibition could be a consequence of Ca2+-Mg2+ competition, Ca . ATP-Mg . ATP competition or a combination of both mechanisms. In the presence of Na+ and Mg2+, Ca2+ inhibited the K+-dependent dephosphorylation of the phosphoenzyme formed from ATP, had no effect on the dephosphorylation in the absence of K+ and inhibited the rephosphorylation of the enzyme. In addition, the steady-state levels of phosphoenzyme were reduced in the presence both of NaCl and of NaCl plus KCl. With 3 mM ATP, Ca2+ alone sustained no more than 2% of the (Na+ + K+)-ATPase activity and about 23% of the Na+-ATPase activity observed with Mg2+ and no Ca2+. With 0.003 mM ATP, Ca2+ was able to maintain about 40% of the (Na+ + K+)-ATPase activity and 27% of the Na+-ATPase activity seen in the presence of Mg2+ alone. However, the E2(K)-E1K conformational change did not seem to be affected. Ca2+ inhibition of the K+-dependent rho-nitrophenylphosphatase activity of the (Na+ + K+)-ATPase followed competition kinetics between Ca2+ and Mg2+. In the presence of 10 mM NaCl and 0.75 mM KCl, the fractional inhibition of the K+-dependent rho-nitrophenylphosphatase activity as a function of Ca2+ concentration was the same with and without ATP, suggesting that Ca2+ indeed plays the important role in this process. In the absence of Mg2+, Ca2+ was unable to sustain any detectable ouabain-sensitive phosphatase activity, either with rho-nitrophenylphosphate or with acetyl phosphate as substrate. 相似文献
19.
General properties of ouabain-sensitive K+ binding to purified Na+,K+-ATPase [EC 3.6.1.3] were studied by a centrifugation method with 42K+. 1) The affinity for K+ was constant at pH values higher than 6.4, and decreased at pH values lower than 6.4. 2) Mg2+ competitively inhibited the K+ binding. The dissociation constant (Kd) for Mg2+ of the enzyme was estimated to be about 1 mM, and the ratio of Kd for Mg2+ to Kd for K+ was 120 : 1. The order of inhibitory efficiency of divalent cations toward the K+ binding was Ba2+ congruent to Ca2+ greater than Zn2+ congruent to Mn2+ greater than Sr2+ greater than Co2+ greater than Ni2+ greater than Mg2+. 3) The order of displacement efficiency of monovalent cations toward the K+ binding in the presence or absence of Mg2+ was Tl+ greater than Rb+ greater than or equal to (K+) greater than NH4+ greater than or equal to Cs+ greater than Na+ greater than Li+. The inhibition patterns of Na+ and Li+ were different from those of other monovalent cations, which competitively inhibited the K+ binding. 4) The K+ binding was not influenced by different anions, such as Cl-, SO4(2-), NO3-, acetate, and glycylglycine, which were used for preparing imidazole buffers. 5) Gramicidin D and valinomycin did not affect the K+ binding, though the former (10 micrograms/ml) inhibited the Na+,K+-ATPase activity by about half. Among various inhibitors of the ATPase, 0.1 mM p-chloromercuribenzoate and 0.1 mM tri-n-butyltin chloride completely inhibited the K+ binding. Oligomycin (10 micrograms/ml) and 10 mM N-ethylmaleimide had no effect on the K+ binding. In the presence of Na+, however, oligomycin decreased the K+ binding by increasing the inhibitory effect of Na+, whether Mg2+ was present or not. 6) ATP, adenylylimido diphosphate and ADP each at 0.2 mM decreased the K+ binding to about one-fourth of the original level at 10 microM K+ without MgCl2 and at 60 microM K+ with 5 mM MgCl2. On the other hand, AMP, Pi, and p-nitrophenylphosphate each at 0.2 mM had little effect on the K+ binding. 相似文献
20.
Cadmium-resistant mutant of Bacillus subtilis 168 with reduced cadmium transport. 总被引:9,自引:2,他引:7
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Cd2+ and Mn2+ accumulation was studied with wild-type Bacillus subtilis 168 and a Cd2+-resistant mutant. After 5 min of incubation in the presence of 0.1 microM 109Cd2+ or 54Mn2+, both strains accumulated comparable amounts of 54Mn2+, while the sensitive cells accumulated three times more 109Cd2+ than the Cd2+-resistant cells did. Both 54Mn2+ and 109Cd2+ uptake, which apparently occur by the same transport system, demonstrated cation specificity; 20 microM Mn2+ or Cd2+ (but not Zn2+) inhibited the uptake of 0.1 microM 109Cd2+ or 54Mn2+. 54Mn2+ and 109Cd2+ uptake was energy dependent and temperature sensitive, but 109Cd2+ uptake in the Cd2+-resistant strain was only partially inhibited by an uncoupler or by a decrease in temperature. 109Cd2+ uptake in the sensitive strain followed Michaelis-Menten kinetics with a Km of 1.8 microM Cd2+ and a Vmax of 1.5 mumol/min X g (dry weight); 109Cd2+ uptake in the Cd2+-resistant strain was not saturable. The apparent Km value for the saturable component of 109Cd2+ uptake by the Cd2+-resistant strain was very similar to that of the sensitive strain, but the Vmax was 25 times lower than the Vmax for the sensitive strain. The Km and Vmax for 54Mn2+ uptake by both strains were very similar. Cd2+ inhibition of 54Mn2+ uptake had an apparent Ki of 3.4 and 21.5 microM Cd2+ for the sensitive and Cd2+-resistant strains, respectively. Mn2+ had an apparent Ki of 1.2 microM Mn2+ for inhibition of 109Cd2+ uptake by the sensitive strain, but the Cd2+-resistant strain had no defined Ki value for inhibition of Cd2+ uptake by Mn2+. 相似文献