低温,高pH胁迫对水稻幼苗根系质膜,液泡膜ATP酶活性的影响

以耐冷性不同的两个水稻品种为材料,比较研究了幼苗根系质膜、液泡膜ＡＴＰ酶对低温（８℃）及高ｐＨ（８．０）胁迫的反应。结果表明：水稻根细胞质膜和液泡膜上均存在Ｃａ＾２＋－ＡＴＰ酶,但活性远低于Ｈ＾＋－ＡＴＰ酶。耐冷品种武育粳３号经低温（８℃）处理２ｄ,根系质膜和液泡膜Ｈ＾２＋－ＡＴＰ酶、Ｃａ＾２＋－ＡＴＰ酶海性均明显升高,至冷处理１２ｄ,Ｈ＾＋－ＡＴＰ酶、Ｃａ＾２＋－ＡＴＰ酶活性有所下降,但仍与对照     

苹果果肉质膜微囊主动动输Ca^2＋的Ca^2＋—ATP酶特性

应用＾４５Ｃａ＾２＋示踪法研究了苹果果肉质膜微囊依赖于Ｃａ＾２＋的ＡＴＰ酶（Ｃａ＾２＋－ＡＴＰ酶）活性与Ｃａ＾２运输之间的关系及激素对该酶活性的影响。结果表明：Ｃａ＾２＋－ＡＴＰ酶存在于质膜上并受载体Ａ２３８７刺激而活性增加,该酶活性与依赖于ＡＴＰ的Ｃａ＾２＋运输依抑制剂ＥＢ,游离Ｃａ＾２＋和ＡＴＰ浓度的变化并呈极为相似的饱和动力学特征,而其ＥＢ半抑制浓度,Ｃａ＾２＋和ＡＴＰ半饱和浓度分别为０．１     

铝胁迫对小麦根系液泡膜ATP酶,焦磷酸酶活性和膜脂组成的效应

研究了铝胁迫下耐铝性不同的两个小麦品种根细胞液泡膜ＡＴＰ酶、焦磷酸酶活性和膜脂的变化。与对照相比,经２０和１００ｍｏｌ／Ｌ的ＡｌＣｌ３处理后,耐铝品种Ａｌｔａｓ６６的液泡膜Ｈ＾＋－ＡＴＰ和和Ｃａ＾２＋－ＡＴＰ酶活性迅速下降;铝敏感品种Ｓｃｏｕｔ６６液泡膜Ｈ＾＋－ＡＴＰ酶和Ｃａ＾２＋－ＡＴＰ酶活性则在２０μｍｏｌ／Ｌ时增加,１００μｍｏｌ／Ｌ时下降。焦磷酸酶活性在Ａｌ－ｔａｓ６６中下降,在Ｓｃｏｕｔ     

热应激大鼠心肌钙代谢的变化及其机理探索

心肌细胞内钙离子对心功能的调节有着极其重要的作用。本研究观察了不同热应激强度下大鼠心肌细胞内质网、线粒体中钙含量,Ｃａ２＋－ＡＴＰ酶活力,内质网Ｃａ２＋主动转运速率及心肌ＡＴＰ含量的变化。研究结果表明,热应激大鼠心肌细胞内质网、线粒体钙含量随肛温升高显著下降;大鼠肛温达４２℃时,其钙含量分别较对照下降３２．２％和４６．５％;心肌细胞内质网和线粒体Ｃａ２＋－ＡＴＰ酶活力亦明显降低,线粒体Ｃａ２＋－ＡＴＰ酶活力下降幅度更为激烈。热应激大鼠心肌内质网Ｃａ２＋主动转运速率和Ｃａ２＋－ＡＴＰ酶活力变化趋势相同,且两者呈密切相关关系。热应激大鼠心肌ＡＴＰ含量亦随肛温升高大幅度降低,当动物肛温超过４２℃时,其可降至对照动物的３７．５％;热应激大鼠心肌ＡＴＰ含量的变化与内质网Ｃａ２＋－ＡＴＰ酶活力关系密切。实验结果提示：热应激心肌细胞质膜受损所致细胞Ｃａ２＋主动转运功能紊乱及心肌能量代谢障碍是心肌钙稳态失调的主要原因;心肌细胞钙代谢紊乱是诱导心肌细胞严重受损,导致热应激机体心血管功能失调,甚至心功能衰竭的重要机制     

铝和铝＋钙对小麦幼苗根尖质膜,液泡膜微囊ATP酶和膜流动性？…

研究了铝和铝＋＿钙对小麦功苗根尖质膜、液泡膜微囊Ｈ＾＋－ＡＴＰ酶、Ｃａ＾２＋－ＡＴＰ酶、Ｍｇ＾２＋－ＡＴＰ酶活性及共动力学参数和膜流动性的影响。在质膜和液泡膜微囊制剂中加入１．０ｍｍｏｌ／Ｌ的ＡＩ＾３＋（ＡＩＣＩ３）时,Ｈ＾２＋－ＡＴＰ囊制剂中加入１．０ｍｍｏｌ／Ｌ的ＡＩ＾３＋（ＡＩＣＩ３）时,Ｈ＾＋－ＡＴＰ酶、Ｃａ＾２＋－ＡＴＰ酶、Ｍｇ＾２＋－ＡＴＰ酶活笥和酶促反应的Ｖｍａｘ及膜流动性下降,而酶     

热暴露大鼠心肌细胞钙稳态及其调节机制的研究

本文观察了离体成年大鼠心室肌肌质网、线粒体Ｃａ＾２＋－ＡＴＰ酶活性和总钙含是到及原代培养乳腺心肌细胞＾４６Ｃａ摄取、活性钙调素相对含量、胞浆内游离钙浓度在不同温度热暴露４０ｍｉｎ后的变化。结果表明：热暴露后,心肌奖浆、肌质网及线粒体中的Ｃａ＾２＋－ＡＴＰ酶活发表 所下降,心肌肌质网及线粒体中的总钙含量亦有降低趋势,心肌细胞活性钙调素相对含量显著下降,＾４６Ｃａ摄取量及胞浆内游离浓度显著升高。提示,     

抗寒剂CR—4对冬小麦幼苗质膜钙泵（Ca^2＋—ATPase）的稳定作用

通过磷酸铈沉淀的细胞化学观察揭示,常温下生长的冬小麦幼苗的Ｃａ２＋ＡＴＰ酶活性主要定位在质膜上,同时,水浸种和抗寒剂浸种的小麦质膜Ｃａ２＋ＡＴＰ酶活性没有差异。然而,小麦幼苗经－７℃冰冻处理１２小时和２４小时后,则表现明显的区别：水浸种的小麦幼苗质膜Ｃａ２＋ＡＴＰ酶活性明显下降,直至完全失活,细胞的精细结构也同时被破坏;而经抗寒剂浸种的小麦幼苗质膜Ｃａ２＋ＡＴＰ酶仍维持较高的活性,细胞结构也保持完整,显示抗寒剂对质膜Ｃａ２＋ＡＴＰａｓｅ酶起着明显的稳定作用。     

苹果果肉质膜微囊主动运输Ca2+的Ca2+-ATP酶特性

应用４５Ｃａ２ ＋ 示踪法研究了苹果果肉质膜微囊依赖于Ｃａ２＋ 的ＡＴＰ 酶（Ｃａ２＋ＡＴＰ酶）活性与Ｃａ２＋ 运输之间的关系及激素对该酶活性的影响。结果表明：Ｃａ２ ＋ＡＴＰ 酶存在于质膜上并受载体Ａ２３１８７ 刺激而活性增加,该酶活性与依赖于ＡＴＰ 的Ｃａ２ ＋ 运输依抑制剂ＥＢ、游离Ｃａ２＋ 和ＡＴＰ浓度的变化并呈极为相似的饱和动力学特征;而其ＥＢ 半抑制浓度,Ｃａ２＋ 和ＡＴＰ 半饱和浓度分别为０ ．１ ,０ ．１ 和５０ μｍｏｌ／Ｌ,从而证实了正是Ｃａ２＋ＡＴＰ酶推动苹果果肉质膜微囊的Ｃａ２＋ 的主动运输。生长素与萘乙酸均可促进苹果果肉质膜微囊Ｃａ２＋ＡＴＰ酶活性和Ｃａ２＋ 吸收,而赤霉素则无此作用。     

神经革苷脂GM3对肌质网Ca^2＋—ATP酶活力的调节

研究了神经节苷脂ＧＭ３参入肌质网膜后Ｃａ＾２＋－ＡＴＰ酶活力的变化。结果表明：ＧＭ３参入肌质网膜后,对肌质网Ｃａ＾２＋－ＡＴＰ酶活性（ＡＴＰ水解活力与转运活力）有明显的激活作用。当参入的ＧＭ３浓度为８μｍｏｌ／Ｌ、参入时间为１２０ｍｉｎ、温度为３０℃时,对Ｃａ＾２＋－ＡＴＰ酶的激活作用最大。     

锌7—金属硫蛋白与镉7—金属硫蛋白抗羟自由基保护线粒 …

制备了Ｚｎ７－与Ｃｄ７－金属硫蛋白。分离出大鼠心肌线粒体。用电子自旋共振自旋标记方法测定线粒体膜脂流动性及膜蛋白构象,运动性,分析了线粒体Ｃａ＾２＋－Ｍｇ６２＋－ＭＡＴＰ酶活性及＾４５Ｃａ摄入。羟自由基损伤使线粒体膜脂流动性下降,膜蛋白构象改变及运动性降低,线粒体Ｃａ＾２＋－Ｍｇ＾２＋ＡＴＰ酶活性降低及＾４５Ｃａ的摄入活性下降。     

Adaptative Responses of Leaf Cells of Rice Seedling to Horizontal Clinostatic Conditions

The authors have studied enzyme cytochemistry, electron probe and ultrastructures in leaf cells of rice seedlings treated under horizontal clinostatic conditions simulating microgravity for 14 days. After treatment, the activity of Ca2+-ATPase in the leaf cell membrane disappeared; the density of total Ca2+ in plasm increased as that density of the cytoskeleton in the leaf cells became sparse; the cell wall was irregular being either or convex in appearance; the structure of chloroplast granas and mitochondria cristae were changed. The authors have proposed the following sequence of events: firstly, the activity of Ca2+-ATPase in leaf cell membrane disappeared; then the calcium-influx in the membrane ceased functioning thereby abolished the cross-membrane Ca2+ gradient; this weakened the assembly of the microtubule and microfilaments, loosened the cytoskeleton, disoriented the movement of the secretory vacuoles, and lead to cell wall thinning and irregularity.     

枸杞体细胞胚发生中Ca2+和ATPase的超微结构定位研究

研究2,4-D诱导枸杞体细胞胚发生中的作用及其与Ca~(2+)含量和ATPase活性时空分布动态之间的关系,以探讨2,4-D诱导植物体细胞胚发生的作用机理。采用超微细胞化学定位的方法,跟踪分析了体细胞胚发生与发育的不同时期,Ca~(2+)和ATPase活性的时空分布动态。结果表明:2,4-D是诱导离体培养的枸杞体细胞进入胚胎状态的关键激素。在含有2,4-D和不含2,4-D的培养条件下,分别诱导枸杞体细胞脱分化后,再转入除去2,4-D的MS培养基上,进行分化培养,结果前者可分化形成体细胞胚,因而称为胚性愈伤组织。后者在相同条件却不能分化形成胚,故称为非胚性愈伤组织。在2,4-D诱导枸杞的胚性愈伤组织中,胚性细胞分化早期的细胞间隙和细胞壁上均有Ca~(2+)沉淀。随着胚性细胞的分化、分裂和多细胞原胚形成,这时Ca~(2+)在细胞内的分布主要集中在细胞膜和液泡膜上;球形胚期在细胞核中Ca~(2+)呈弥散性分布。在此过程中,ATPase活性时空分布与Ca~(2+)的定位变化具有高度一致性,仅仅稍滞后于Ca~(2+)出现的时间。而在胚性细胞分化早期,ATPase活性同样位于质膜上,随后在液泡和细胞核都可见ATPase活性分布。而在非胚性愈伤组织中,则未见Ca~(2+)和ATPase活性呈时空动态分布,而且随着非胚性细胞的液泡化,无论是Ca~(2+)含量,还是ATPase活性都呈逐渐降低的趋势。表明Ca~(2+)和ATPase活性变化与2,4-D诱导的胚性细胞分化和发育密切相关。并由此推测,Ca~(2+)和ATPase的时空分布对胚性细胞分化中的信息传递和调控相关基因表达起着关键性作用。     

枸杞体细胞胚发生中Ca^2＋和ATPase的超微结构定位研究

研究2,4-D诱导枸杞体细胞胚发生中的作用及其与Ca^2 含量和ATPase活性时空分布动态之间的关系,以探讨2,4-D诱导植物体细胞胚发生的作用机理。采用超微细胞化学定位的方法,跟踪分析了体细胞胚发生与发育的不同时期,Ca^2 和ATPase活性的时空分布动态。结果表明：2,4-D是诱导离体培养的枸杞体细胞进入胚胎状态的关键激素。在含有2,4-D和不含2,4-D的培养条件下,分别诱导枸杞体细胞脱分化后,再转入除去2,4-D的MS培养基上,进行分化培养,结果前者可分化形成体细胞胚,因而称为胚性愈伤组织。后者在相同条件却不能分化形成胚,故称为非胚性愈伤组织。在2,4-D诱导枸杞的胚性愈伤组织中,胚性细胞分化早期的细胞间隙和细胞壁上均有Ca^2 沉淀。随着胚性细胞的分化、分裂和多细胞原胚形成,这时Ca^2 在细胞内的分布主要集中在细胞膜和液泡膜上;球形胚期在细胞核中Ca^2 呈弥散性分布。在此过程中,ATPase活性时空分布与Ca^2 的定位变化具有高度一致性,仅仅稍滞后于Ca^2 出现的时间。而在胚性细胞分化早期,ATPase活性同样位于质膜上,随后在液泡和细胞核都可见ATPase活性分布。而在非胚性愈伤组织中,则未见Ca^2 和ATPase活性呈时空动态分布,而且随着非胚性细胞的液泡化,无论是Ca^2 含量,还是ATPase活性都呈逐渐降低的趋势。表明Ca^2 和ATPase活性变化与2,4-D诱导的胚性细胞分化和发育密切相关。并由此推测,Ca^2 和ATPase的时空分布对胚性细胞分化中的信息传递和调控相关基因表达起着关键性作用。     

花生不同生育期小叶脉转移细胞形态变化和ATP酶活性定位的研究

迄今已在上千种高等植物中看到具有胞壁内突生长的转移细胞,认为它是一种输送溶质的细胞,在源库二端行使光合产物的垭距离运输。转移细胞质膜上具有较强的ATP酶活性,在发育成熟的转移细胞质膜上ATP酶活     

枸杞胚性细胞分化的超微结构和ATP酶的细胞化学定位研究

枸杞的胚性细胞多由愈伤组织表层的薄壁细胞分化而来,与愈伤组织中未分化的细胞相比,胚性细胞呈卵圆形,细胞核大,核仁明显,细胞质浓厚并含有丰富的细胞器,细胞壁较薄,细胞间有胞间连丝相通;胚性细胞发育到晚期细胞壁加厚,胞间连丝逐渐消失,细胞核向一端偏移,有大液泡形成;胚性细胞的第一次分裂多为均等分裂,形成二细胞原胚,继续分裂形成多细胞原胚;组成多细胞原胚胚体的细胞核大,核形状不规则,细胞质浓厚,细胞器丰富,在质体中出现淀粉的积累。在胚性细胞发育的早期,ATP酶活性主要位于质膜上,随后在液泡内和细胞核中都出现ATP酶活性的分布;随着胚性细胞壁的加厚,细胞壁加厚处和细胞间隙中也出现ATP酶活性反应;当多细胞原胚形成后,ATP酶活性反应主要定位于液泡膜上。由此分析了结构特征、ATP酶活性定位变化与胚性细胞分化的关系。     

A high affinity, calmodulin-responsive (Ca2+ + Mg2+)-ATPase in isolated bone cells

Although acute alterations in Ca2+ fluxes may mediate the skeletal responses to certain humoral agents, the processes subserving those fluxes are not well understood. We have sought evidence for Ca2+-dependent ATPase activity in isolated osteoblast-like cells maintained in primary culture. Two Ca2+-dependent ATPase components were found in a plasma membrane fraction: a high affinity component (half-saturation constant for Ca2+ of 280 nM, Vmax of 13.5 nmol/mg per min) and a low affinity component, which was in reality a divalent cation ATPase, since Mg2+ could replace Ca2+ without loss of activity. The high affinity component exhibited a pH optimum of 7.2 and required Mg2+ for full activity. It was unaffected by potassium or sodium chloride, ouabain or sodium azide, but was inhibited by lanthanum and by the calmodulin antagonist trifluoperazine. This component was prevalent in a subcellular fraction which was also enriched in 5'-nucleotidase and adenylate cyclase activities, suggesting the plasma membrane as its principal location. Osteosarcoma cells, known to resemble osteoblasts in their biological characteristics and responses to bone-seeking hormones, contained similar ATPase activities. Inclusion of purified calmodulin in the assay system caused small non-reproducible increases in the Ca2+-dependent ATPase activity of EGTA-washed membranes. Marked, consistent calmodulin stimulation was demonstrated in membranes exposed previously to trifluoperazine and then washed in trifluoperazine-free buffer. These results indicate the presence of a high affinity, calmodulin-sensitive Ca2+-dependent ATPase in osteoblast-like bone cells. As one determinant of Ca2+ fluxes in bone cells, this enzyme may participate in the hormonal regulation of bone cell function.     

A cytochemical and immunocytochemical analysis of the wall labyrinth apparatus in leaf transfer cells in Elodea canadensis

Background and Aims

Transfer cells are plant cells specialized in apoplast/symplast transport and characterized by a distinctive wall labyrinth apparatus. The molecular architecture and biochemistry of the labyrinth apparatus are poorly known. The leaf lamina in the aquatic angiosperm Elodea canadensis consists of only two cell layers, with the abaxial cells developing as transfer cells. The present study investigated biochemical properties of wall ingrowths and associated plasmalemma in these cells.

Methods

Leaves of Elodea were examined by light and electron microscopy and ATPase activity was localized cytochemically. Immunogold electron microscopy was employed to localize carbohydrate epitopes associated with major cell wall polysaccharides and glycoproteins.

Key Results

The plasmalemma associated with the wall labyrinth is strongly enriched in light-dependent ATPase activity. The wall ingrowths and an underlying wall layer share an LM11 epitope probably associated with glucuronoarabinoxylan and a CCRC-M7 epitope typically associated with rhamnogalacturonan I. No labelling was observed with LM10, an antibody that recognizes low-substituted and unsubstituted xylan, a polysaccharide consistently associated with secondary cell walls. The JIM5 and JIM7 epitopes, associated with homogalacturonan with different degrees of methylation, appear to be absent in the wall labyrinth but present in the rest of cell walls.

Conclusions

The wall labyrinth apparatus of leaf transfer cells in Elodea is a specialized structure with distinctive biochemical properties. The high level of light-dependent ATPase activity in the plasmalemma lining the wall labyrinth is consistent with a formerly suggested role of leaf transfer cells in enhancing inorganic carbon inflow. The wall labyrinth is a part of the primary cell wall. The discovery that the wall ingrowths in Elodea have an antibody-binding pattern divergent, in part, from that of the rest of cell wall suggests that their carbohydrate composition is modulated in relation to transfer cell functioning.     

Mimosa pudica adenosine triphosphatase]

The morphological structure (pulvinus P1, P2 and P3) directly involved in the seismonastic movements of the Mimosa pudica leaf have been used to isolate: 1) "soluble" ATPase, loosely bound to pulvinus structures; 2) Ca, Mg-dependent ATPase, which is tightly bound to pulvinus structures and is extracted by a saline solution of high ionic strength, used to isolate actomyosin from muscles and non-muscle motile cells; 3) ATPase bound to the pulvinus membrane structures, which is solubilized by the detergents, e. g. Triton X-100 and Tween-80, and is similar to membrane ATPase. Physico-chemical and kinetic studies of the APSases have shown that Ca,Mg-ATPase is similar to the ATPases from muscle and non-muscle motile cells in a number of characteristics, e. g. solubility in saline solution of high ionic strength, aggregability in a solution of lower ionic strength, activation by bivalent metal ions, pH-optimum, specificity for substrates, etc. The protein composition of the ATPases has been determined by gel-electrophoresis in polyacrylamide gel. The molecular weight of purified Ca,Mg-ATPase from Mimosa pudica pulvinus is found to be 139 000. The role of ATPases in seismonastic movements of the Mimosa pudica leaf is discussed.     

一种特殊的长寿细胞:毛竹茎秆纤维细胞

利用显微和细胞化学方法,对毛竹(Phyllostachys edulis)茎秆纤维次生壁形成过程中超微结构变化以及ATP酶、Ca2 -ATPase和酸性磷酸酶的超微细胞化学定位进行了研究.研究发现,次生壁形成早期,细胞核具有双层核膜,染色质凝聚,可见大量的线粒体、粗面内质网和高尔基体等细胞器存在于纤维细胞中;随后,双层核膜消失,细胞器将逐渐解体,多泡体开始出现在纤维细胞的细胞质;随着年龄的增加,纤维细胞壁逐渐增厚,并出现多层结构现象,而运输小泡、细胞膜、胞间连丝和凝聚的染色质将持续存在.在次生壁形成的整个过程中,ATP酶、Ca2 -ATPase和酸性磷酸酶在运输小泡、细胞膜、质膜内陷、胞间连丝和凝聚的染色质中将持续存在.结果表明,毛竹茎秆纤维细胞是一种不同于木本双子叶植物的长寿细胞,纤维原生质体中ATP酶和酸性磷酸酶的持续存在与次生壁的持续增厚密切相关.