首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 15 毫秒
1.
The cold inhibited functions of skin thermoreceptors, of the thermoregulation centre, and the respiration centre during deep hypothermia can be restored without rewarming the body. The methods used were developed to test the hypothesis that during deep hypothermia calcium ion concentration [Ca2+]i in the cytoplasm increases. This causes a perturbation of cell metabolism, the impairment of cell membrane function that cause the inhibition of cell functioning, resulting in cell death. Such an increase in [Ca2+]i most likely would result from an energy deficit in a deeply cooled cell, which would compromise the processes that maintain the [Ca2+]i at about 10−7 M. These processes require large amounts of energy since they occur against a large concentration gradient. With the use of EDTA the extracellular concentration of Ca2+ has been lowered by 15–27%, so reducing the concentration gradient for Ca2+ between the cell and the medium and in consequence facilitated the process the extrusion of cell Ca2+.

During a period of cooling, sufficient to impair normal functioning, the experimental lowering of blood Ca2+ allowed the restoration of normal function without the need to rewarm. In such cases the animals survived after cooling the body to temperatures at which they would normally have succumbed. The data presented support the stated hypothesis that the impairment of cellular function in mammals by low temperatures is the result of an uncorrected rise in [Ca2+]i.  相似文献   


2.
The amylases produced by a Bacillus stearothermophilus were purified through a series of four steps. Two separable enzyme fractions having starch hydrolysing activity were eluted from a DEAE-cellulose column by NaCl gradient elution. The homogeneity of the purified enzymes was checked on polyacrylamide gel electrophoresis. The product formation studies indicated that fraction I was an -amylase whereas fraction II was a β-amylase. The molecular weights were determined to be 48 000 and 57 000 and the carbohydrate moiety was found to be 13.2 and 0.8% for - and β-amylase, respectively. The protein digest of these enzymes indicated a total number of 15 amino acids with aspartic and glutamic acid showing the highest value. The purified amylase showed maximal activity at 80°C and pH 6.9. Fe3+, Cd2+, Pb2+, Hg2+, Ni2+ and Ag1+ were potent inhibitors whereas Zn2+, Mg2+, Mn2+ and Al3+ were mild inhibitors. Ca2+, Ba2+, Sr2+ and K+ stimulated amylase activity in the order of Ca2+ > Ba2+ > Sr2+ > K+. PCMB, EDTA and sodium iodoacetate were inhibitory whereas glutathione (GSH) and cysteine afforded protection of enzyme activity. EDTA showed dose-dependent noncompetitive inhibition of both - as well as β-amylase activities. EDTA inhibition was reversed by the addition of Ca2+ and PCMB inhibition by the addition of glutathione (reduced). The Km for - and β-amylases were found to be 1.05 and 1.25 mg starch per ml, respectively.  相似文献   

3.
The effects of calcium ions (Ca2+) on the stability of artichoke (Cynara scolymus L.) peroxidase (AKPC) have been studied. The thermal stability of AKPC was improved by the addition of Ca2+; the melting temperature increased by 20 °C and the deactivation energy by 26 kJ mol−1. AKPC was stable in a selection of organic solvents but was less active with 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) than under aqueous conditions. Ca2+-free AKPC retained more activity in the presence of organic solvents due to its better maintenance of the rate of compound I formation with hydrogen peroxide (H2O2) compared to AKPC-Ca2+. AKPC retained at least 75% activity over 24 h in the pH range 3.0–10.5 and about 50% over 1 month at pH 7.0 or 5.5, irrespective of the Ca2+ content. AKPC-Ca2+ was considerably more resistant to inactivation by H2O2 than Ca2+-free AKPC suggesting that the presence of Ca2+ boosts turnover under oxidizing conditions. AKPC has been applied as an alternative to horseradish peroxidase (HRP) in glucose concentration assays; the presence of Ca2+ or of the Ca2+ chelating agent ethylenediaminetetraacetic acid made no difference to the final result. The possibility is discussed that addition and removal of a labile Ca2+ from AKPC could be used to control enzyme activity both in vivo and in vitro.  相似文献   

4.
An alkaline protease produced from a Bacillus sp. was stimulated by the metal ions Ca2+, Mg2+ and Mn2+; with Ca2+ having the maximum effect. The thermal stability of the enzyme was also enhanced to varying degrees in the presence of these ions.  相似文献   

5.
Pseudomonas aeruginosa PD100 capable of producing an extracellular protease was isolated from the soil collected from local area (garbage site) from Shivage market in Pune, India. The purified protease showed a single band on native and SDS-PAGE with a molecular weight of 36 kDa on SDS-PAGE. The optimum pH value and temperature range were found to be 8 and 55–60 °C, respectively. The enzyme exhibited broad range of substrate specificity with higher activity for collagen. The enzyme was inhibited with low concentration of Ag2+, Ni2+, and Cu2+. β-Mercaptoethanol was able to inactivate the enzyme at 2.5 mM, suggesting that disulfide bond(s) play a critical role in the enzyme activity. Studies with inhibitors showed that different classes of protease inhibitors, known to inhibit specific proteases, could not inhibit the activity of this protease. Amino acid modification studies data and pKa values showed that Cys, His and Trp were involved in the protease activity. P. aeruginosa PD100 produces one form of protease with some different properties as compared to other reported proteases from P. aeruginosa strains. With respect to properties of the purified protease such as pH optimum, temperature stability with capability to degrade different proteins, high stability in the presences of detergents and chemicals, and metal ions independency, suggesting that it has great potential for different applications.  相似文献   

6.
To study the mechanism of action of diflubenzuron (DFB) and other benzoylphenylureas, we have initially hypothesized that their action may be related to exocytosis: to test the hypothesis, we obtained an intracellular vesicle preparation from the homogenate of integument of newly molted American cockroachs (Periplaneta americana L.) in 10 mM MES buffer containing 250 mM sucrose (isotonic) and 2.5 mM MgSO4, at pH 6.6. By studying DFB's effect on various ion transporting activities, we demonstrated that calcium uptake in this intracellular particulate preparation was significantly inhibited by DFB at low concentrations (e.g., 10−8 M). Such an inhibitory effect of DFB on Ca2+ uptake was eliminated by the addition of ionophores or membrane disruptors, as well as the sonication of vesicle preparation. On the other hand, oligomycin, protein phosphorylation modulators, Na+, and Li+ did not affect the calcium uptake. Among ionophores, agents disrupting H+ gradients (e.g. FCCP and NEM) totally eliminated 45Ca uptaking activity by vesicles as well as the inhibitory effect of DFB. Among calcium ion modulators, calmodulin inhibitors such as calmidazolium and trifluoperazine decreased the Ca2+-uptake, whereas membrane calcium channel blocker, verapamil, did not. ATP and γ-S-GTP stimulated Ca2+ uptake. However, the former increased only the DFB insensitive portion and the latter largely the DFB sensitive part of Ca2+. Together these data support the hypothesis that the action site of DFB in this preparation is the GTP-dependent Ca2+ transport process which is coupled to vacuolar type intracellular vesicles in the integument cells.  相似文献   

7.
为探讨大黄鱼幼鱼在低氧及酸化胁迫下机体离子调节情况,本研究探讨了低氧(溶解氧量DO 3.5 mg·L-1,pH 8.1)、酸化(DO 7.0 mg·L-1,pH 7.35)以及低氧酸化协同胁迫(DO3.5 mg·L-1,pH 7.35)对大黄鱼幼鱼鳃组织结构以及离子调节相关生理指标的影响.结果 表明:低氧胁迫下,大黄鱼...  相似文献   

8.

1. 1. (Mg2+ + Ca2+) ATPases of microsomal and synaptic membrane preparations from immature and adult rat brain were activated by calcium (0.1–10 μM), maximal activation was found at 3 μM. The increase in (Mg2+ + Ca2+) ATPase seen during development was greatest in the synaptic membrane preparations.

2. 2. At 37°C both Na+ or K+ at concentrations higher than 30 mM inhibited the microsomal Mg2+ ATPase, but the (Mg2+ + Ca2+) ATPase was stimulated by both Na+ and K+. Synaptic membrane Mg2+ ATPase was inhibited by concentrations higher than 100 mM K+; Na+ however stimulated this enzyme at all concentrations. Much of this Na+ stimulated activity was ouabain sensitive. Synaptic membrane (Mg2+ + Ca2+) ATPase was stimulated by Na+ or K+, this stimulation follows approximate saturation kinetics with an apparent Km of 18.8 mM Na+ or K+.

3. 3. Arrhenius plots of microsomal (Mg2+ + Ca2+) ATPase were curvilinear, but two intersecting lines with a break at 20°C could be fitted. The calculated energies of activation from these lines were very similar in immature and adult preparations. The synaptic membrane preparation (adult) also gave a curvilinear plot; but two intersecting lines with a break at 25°C could be fitted to the data. These lines had slopes of 21 and 28 Kcal mole−1 above and below the break, respectively. The immature preparation when made using EDTA gave a Arrhenius plot of very similar form to the adult preparation. Without EDTA however the Arrhenius plot was complex with a plateau at 25–32°C. Pretreatment with EDTA activated the synaptic membrane (Mg2+ + Ca2+) ATPase from both immature and adult brain.

Author Keywords: Brain; ATPase; temperature; development; synaptic membranes  相似文献   


9.
Treatment of the oxygen-evolving photosystem II preparation from the thermophilic cyanobacterium Synechococcus sp. with EDTA inhibited electron flow from Z to P680 and consequently induced a back electron flow from Qa to P680+. The inhibition was reversed fully by Ca2+and partially by Mn2+ and Mg2+ when EDTA-treated preparations had been incubated with respective divalent metal cations for several minutes, whereas diphenylcarbazide had no effect on the recombination between qa and P680+ in EDTA-treated preparations. It is concluded that Ca2+ is essential for electron transport from Z to P680.

Oxygen evolution Electron transport Photosystem II Ca2+ Thermophilic cyanobacterium  相似文献   


10.
为探究H2S信号在苜蓿(Medicago sativa)体内调节气孔运动的作用,及在此过程中H2S与Ca2+的关系,以蒺藜苜蓿(Medicago truncatula)的野生型和钙离子转运体突变体为试验材料,分别从转录水平、细胞水平和生理水平开展研究。采用qRT-PCR比较相关基因的表达量变化、荧光探针显示体内Ca2+含量、电极法测定H2S含量、光学显微镜观察和测量气孔孔径等。结果表明:蒺藜苜蓿突变体NF3011和NF2734体内H2S的含量与野生型相比极显著降低(P<0.01);H2S信号在一定程度上抑制钙离子转运体编码基因MTR_6g027580的表达;外源生理浓度H2S熏蒸可诱导蒺藜苜蓿气孔关闭,与Ca2+通道阻断剂LaCl3联合处理对野生型气孔运动未产生影响,而在突变体中的结果截然相反;利用荧光探针测定保卫细胞内的Ca2+含量,所得结果与气孔孔径的变化规律完全一致。综上所述,H2S信号促进叶片保卫细胞内Ca2+的含量增加,最终表现为植物气孔孔径变小,在此过程中胞内Ca2+含量变化主要通过Ca2+转运体进行,少部分依赖Ca2+离子通道。该研究结果不仅在理论上丰富了H2S信号的作用机制,更具应用于苜蓿生产实践并推广于其他作物的潜力。  相似文献   

11.
To elucidate the relationship between intracellular free Ca2+ concentration ([Ca2+]i) and Ca2+-signalling by the sarcoplasmic reticulum (SR) in Ca2+-overloaded heart muscle cells, the direct effects of “basal” [Ca2+]i on calcium waves were investigated by altering the membrane potential. When basal inter-calcium wave (BCW) [Ca2+]i was maintained at a high level, (i) calcium waves showed more gradual and more rapidly suppressed increase in [Ca2+]-profile (P < 0.005), and (ii) calcium waves occurred at a significantly higher frequency and velocity (259% and 137%), than when low BCW [Ca2+]i was maintained. Similar investigations on inhibition of the Na+-Ca2+ exchanger, however, showed that membrane potential did not elicit direct effects on calcium waves. These results showed that the elevation of BCW [Ca2+]i per se directly influences Ca2+-signalling in heart muscle cells through non-equilibrated release-restoration Ca2+-handling by the SR.  相似文献   

12.
为了探明褪黑素(MT)和钙离子(Ca2+)在调控植物耐热性中是否存在互作关系,以黄瓜幼苗为试材,分析了内源MT和Ca2+对高温胁迫的响应;并通过叶面喷施100 μmol·L-1 MT、10 mmol·L-1 CaCl2、3 mmol·L-1乙二醇二乙醚二胺四乙酸(EGTA,Ca2+螯合剂)+100 μmol·L-1 MT、0.05 mmol·L-1氯丙嗪(钙调素拮抗剂,CPZ)+100 μmol·L-1 MT、100 μmol·L-1氯苯丙氨酸(p-CPA,MT合成抑制剂)+10 mmol·L-1 CaCl2和去离子水(H2O),研究高温下(42/32 ℃)外源MT和Ca2+对黄瓜幼苗活性氧积累、抗氧化系统及热激转录因子(HSF)和热激蛋白(HSPs)等的影响。结果表明: 黄瓜幼苗内源MT和Ca2+均受高温胁迫诱导;外源MT可上调常温下钙调素蛋白(CaM)、钙依赖蛋白激酶(CDPK5)、钙调磷酸酶B类蛋白(CBL3)、CBL结合蛋白激酶(CIPK2)mRNA表达;CaCl2处理的MT合成关键基因色氨酸脱羧酶(TDC)、5-羟色胺-N-乙酰转移酶(SNAT)和N-乙酰-5-羟色胺甲基转移酶(ASMT)水平也显著升高,MT含量快速增加。MT和CaCl2可显著增强高温下黄瓜的抗氧化能力,减少活性氧(ROS)积累,同时上调HSF7HSP70.1HSP70.11 mRNA表达,从而减轻高温胁迫引起的过氧化伤害,植株热害症状明显减轻,热害指数和电解质渗漏率显著降低。加入EGTA和CPZ后,MT对黄瓜幼苗抗氧化能力和热激蛋白表达的促进效应明显减弱,Ca2+对高温下黄瓜幼苗过氧化伤害的缓解效应也被p-CPA逆转。可见,MT和Ca2+均可诱导黄瓜幼苗的耐热性,二者在热胁迫信号转导过程中存在互作关系。  相似文献   

13.
The role of Ca2+ in glycerol dissimilation under hypoosmotic stress in the halotolerant alga Dunaliella tertiolecta was investigated using a pharmacological approach. A stretch-activated Ca2+ channel blocker, GdCl3, inhibited glycerol dissimilation under hypoosmotic stress. However, addition of voltage-dependent Ca2+ channel blockers and inhibitors of mitochondrial and endoplasmic reticulum Ca2+ channels did not affect the glycerol dissimilation under hypoosmotic stress. The results of the present study suggest that the influx of Ca2+ from the extracellular space via the stretch-activated Ca2+ channels localized in the plasma membrane is required for the transduction of osmotic signal of D. tertiolecta.  相似文献   

14.
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   

15.
Heparin was fractionated on an affinity column of bovine milk lipoprotein lipase (LpL) immobilized to Affi-Gel-15. The bound heparin, designated high-reactive heparin (HRH), enhanced LpL activity, presumably by stabilizing the enzyme against denaturation. The unbound heparin fraction had no observable effect on the initial rate of enzyme activity. However, at longer times of incubation there was inhibition of LpL activity. LpL-specific HRH also showed a high, Ca2+-dependent precipitating activity towards human plasma low density lipoproteins (LDL). Since LpL and LDL both bind to heparin-like molecules at the surface of the arterial wall, we suggest that their similar heparin-binding specificity may have physiological consequences as it relates to the development of atherosclerosis.

Heparin binding Lipoprotein lipase LDL Apolipoprotein Lipolysis  相似文献   


16.
Vertebrate embryos generate striking Ca2+ patterns, which are unique regulators of dynamic developmental events. In the present study, we used zebrafish embryos as a model system to examine the developmental roles of Ca2+ during gastrulation. We found that gastrula stage embryos maintain a distinct pattern of cytosolic Ca2+ along the dorsal–ventral axis, with higher Ca2+ concentrations in the ventral margin and lower Ca2+ concentrations in the dorsal margin and dorsal forerunner cells. Suppression of the endoplasmic reticulum Ca2+ pump with 0.5 μM thapsigargin elevates cytosolic Ca2+ in all embryonic regions and induces a randomization of laterality in the heart and brain. Affected hearts, visualized in living embryos by a subtractive imaging technique, displayed either a reversal or loss of left–right asymmetry. Brain defects include a left–right reversal of pitx2 expression in the dorsal diencephalon and a left–right reversal of the prominent habenular nucleus in the brain. Embryos are sensitive to inhibition of the endoplasmic reticulum Ca2+ pump during early and mid gastrulation and lose their sensitivity during late gastrulation and early segmentation. Suppression of the endoplasmic reticulum Ca2+ pump during gastrulation inhibits expression of no tail (ntl) and left–right dynein related (lrdr) in the dorsal forerunner cells and affects development of Kupffer’s vesicle, a ciliated organ that generates a counter-clockwise flow of fluid. Previous studies have shown that Ca2+ plays a role in Kupffer’s vesicle function, influencing ciliary motility and translating the vesicle’s counter-clockwise flow into asymmetric patterns of gene expression. The present results suggest that Ca2+ plays an additional role in the formation of Kupffer’s vesicle.  相似文献   

17.
In this paper, the effects of some chemical and physical factors such as temperature, pH values, glycerol, and divalent metal cations on the protease activity of venom from jellyfish, Rhopilema esculentum Kishinouye, were assayed. Protease activity was dependent on temperature and pH values. Zn2+, Mg2+, and Mn2+ in sodium phosphate buffer (0.02 M, pH 8.0) could increase protease activity. Mn2+ had the best effects among the three metal cations and the effect was about 20 times of that of Zn2+ or Mg2+ and its maximal protease activity was 2.3 × 105 U/mL. EDTA could increase protease activity. PMSF had hardly affected protease activity. O-Phenanthroline and glycerol played an important part in inhibiting protease activity and their maximal inhibiting rates were 87.5% and 82.1%, respectively.  相似文献   

18.
以冰叶日中花(Mesembryanthemum crystallinum L.)实生苗为材料,经NaCl、NaCl+ CaCl_2、NaCl+LaCl_3处理后,利用电感耦合等离子发射光谱仪检测叶、茎、根中Na~+、K~+、Ca~(2+)、Mg~(2+)含量,计算K~+/Na~+、Ca~(2+)/Na~+和Mg~(2+)/Na~+比值,利用非损伤微测技术测定根尖Na~+流和K~+流,研究盐胁迫下钙在维持离子平衡中的作用。结果显示,NaCl处理后,冰叶日中花各器官中Na~+含量增加,K~+、Ca~(2+)、Mg~(2+)含量降低,离子比值降低;CaCl_2处理降低了Na~+含量,提高了K~+、Ca~(2+)、Mg~(2+)含量,离子比值升高,而LaCl_3处理后的结果相反。经NaCl处理24 h后,冰叶日中花根尖Na~+和K~+明显外流,加入CaCl_2后,Na~+外流速度显著增加,K~+外流速度受到抑制,而加入LaCl_3后则降低了Na~+的外流速度,促进了K~+的外流。研究结果表明冰叶日中花受到盐胁迫后,钙参与了促进根部Na~+外排、抑制K~+外流的过程,进而保持各器官中较低的Na~+含量,表明钙在维持和调控离子平衡中起到重要作用。  相似文献   

19.
高Ca2+环境对许多植物的生长不利, 因此研究植物对高Ca2+环境的适应机制非常重要。研究发现, 拟南芥(Ara- bidopsis thaliana)镁转运体MGT7功能缺失突变体mgt7-1mgt7-2具有高Ca2+敏感表型: 在高Ca2+培养基上, 相对于野生型Col-0, 突变体叶鲜重显著下降, 但根长无显著差异。高Ca2+MGT7启动子活性和包括MGT7在内的镁转运体基因表达无显著调节作用。Col-0与mgt7突变体之间, 在外加Ca2+诱导细胞质Ca2+瞬时升高和Ca2+含量方面无显著差异; 但是, 在正常和高Ca2+培养基上, mgt7突变体的Mg含量均显著低于Col-0。高Ca2+显著抑制Col-0和mgt7突变体内Mg的积累。因此我们假设, mgt7突变体的高Ca2+敏感表型是由于其体内Mg含量下降导致的。进一步的研究证实, 只有增加培养基中Mg2+的含量, 而不是N、P、K和S, 才可以使突变体的高Ca2+敏感表型得到恢复。  相似文献   

20.
为探究微咸水磁化处理条件下植株的离子稳态特征,以欧美杨I-107一年生扦插苗为试材,于生长季节分别采用Hoagland营养液和4.0 g·L-1 NaCl微咸水,经磁化处理后连续灌溉30 d.采用原子吸收分光光度法对叶片和根系中K+、Na+、Ca2+和Mg2+含量进行测定,分析离子平衡系数(K)和根-叶之间的离子选择性运输系数(SXi,Na).结果表明: 与非盐分胁迫处理相比,盐分胁迫处理根系和叶片中Na+和Ca2+含量及SK,NaSMg,Na升高,K+和Mg2+含量、K+/Na+SCa,Na降低.与非磁化微咸水灌溉处理相比,磁化微咸水灌溉处理的根系和叶片中Na+含量降低、K+含量及K+/Na+提高;根系和叶片中Ca2+含量降低、Mg2+含量提高;磁化微咸水灌溉处理中K提高,且叶片中K值显著高于根系;SK,NaSMg,Na较非磁化微咸水灌溉提高,SCa,Na较其降低.磁化微咸水灌溉中根系和叶片Na+积累量减少,K+、Ca2+和Mg2+含量增加,且维持了较高水平的K+/Na+,这有利于植株整株水平生理代谢的调控.因此,盐分胁迫下磁化作用可通过调节离子的选择性吸收和运输来维持植株体内的离子平衡.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号