植物细胞生物碱跨膜传递模型研究(Ⅱ)-膜电位影响下的饱和特性模型

在中性生物碱饱和特性模型的基础上,引入膜电位的影响,考虑生物碱阳离子通量对总生物碱扩散通量的影响,从而导出膜电位影响的饱和特性模型。将模型在特定参数下积分,对实验结果进行数值模拟,结果表明,所建模型可很好描述各种生物碱的跨膜传递过程,为建立描述生物碱跨膜传递过程的普适性模型提供了理论基础。     

植物细胞胞内生物碱跨膜传递模型研究(Ⅰ)-中性生物碱饱和特性模型

针对制约植物细胞生物碱释放的生物碱跨膜传递这一物理过程,引入有载体参与的主动运输过程来描述植物细胞生物碱的跨膜传递过程,将Ｍｉｃｈａｅｌｉｓ－Ｍｅｎｔｅｎ酶促反应动力学公式应用于载体转运,建立了植物细胞中产物释放的饱和特性模型。将模型在特定参数下进行积分,对已有的实验现象进行数值模拟,并与实验结果比较表明,所建模型能很好地描述低Ｐｋａ值生物碱的各种跨膜传递现象,并能同时呈现生物碱跨膜传递的线性特性和饱和特性,从而为长期对立的生物碱跨膜传递的简单扩散观点和主动运输观点的统一提供了理论依据     

跨膜Ca^2＋梯度对肌质网Ca^2＋—ATP酶调节的特异性

我们曾报道跨膜Ｃａ＾２＋梯度可通过膜脂影响肌质网Ｃａ＾２＋－ＡＴＰ酶的构象和活性。本文就跨膜Ｃａ＾２＋－ＡＴＰ酶的构象和活性。本文就跨膜Ｃａ＾２＋梯度对肌质网Ｃａ＾２＋－ＡＴＰ酶的调节是否具有特异性作进一步研究。结果表明这种特异性表现在两方面：一是跨膜Ｃａ＾２＋梯度对肌质网Ｃａ＾２＋－ＡＴＰ酶功能的调节不能归结于跨膜Ｃａ＾２＋深度梯度所导致的膜电位的作用,离子载体ＦＣＣＰ可消除跨膜电位但并不影响肌     

大鼠海马神经元Na~ Ca~(2 )交换电流在低氧时的变化

目的：探讨在低氧性脑损伤发生过程中,Ｎａ＋Ｃａ２＋ 交换体在细胞内钙超载中的作用。方法：采用全细胞膜片钳方法,在急性分离海马神经元上观察低氧对Ｎａ＋Ｃａ２＋ 交换电流的电流电压（ＩＶ） 曲线的影响。结果：在整个膜电位水平,Ｎａ＋Ｃａ２＋ 交换电流幅值均不同程度的增加,在正膜电位水平呈现一显著的外向电流。１０ ｍＶ 时,电流幅值从（９２ ．８３ ±２０．８）ｐＡ上升到（１３０ ．６７ ±２６．８８）ｐＡ（ Ｐ＜０ ．０５） ,而在５０ ｍ Ｖ,其电流幅值从（－ ７４ ．６７ ±１１ ．８４）ｐＡ上升到（－ ５８ ．５±１０ ．７１）ｐＡ（Ｐ＜ ０ ．０５）。结论：低氧时Ｎａ＋Ｃａ２＋ 交换电流呈外向性,这种改变有利于低氧后通过Ｎａ＋Ｃａ２＋ 交换的外向转运方式排出细胞内钠,并交换钙进入细胞     

大鼠海马神经元Na^2＋—Ca^2＋交流电流在低氧时的变化

目的 探讨在低氧性脑损伤发生过程中,Ｎａ＾２＋－Ｃａ＾２＋交换体在内钙超载中的作用。方法 采用全细胞膜片钳方法,在急性分离海马神经元上观察低氧对Ｎａ＾２＋－Ｃａ＾２＋交换电流的电流－电压（Ｉ－Ｖ）曲线的影响。结果 在整个膜电位水平,Ｂａ＾＋－Ｃａ＾２＋交流电流幅值均不同程度的增加,在正膜电位水平呈现一显著的外向电流。１０ｍｖ时,电流幅值从（９２．８３±２０．８）ｐＡ上升到（１３０．６７±２６．８８     

蜂毒肽在磷脂膜上取向的高效液相色谱和质谱研究

利用脂质体模型系统研究了跨膜电位对蜂毒肽在磷脂磷上取向的影响,采用高效液相色谱与质谱技术相结合方法,分析了蜂毒肽与磷脂膜作用后的胰蛋白酶酶解产物,结果发现,当蜂毒肽分子与没有跨膜电位的脂质体结合后,它的所有酶切位点都能够酶有效切断,当蜂毒肽与带负向膜电全的脂质体结合后,其Ｎ端的一个酶切位点被封闭,从而说明跨膜电位造成了蜂毒肽在膜上的重瓣取向。     

跨膜Ca~(2+)梯差对大豆下胚轴质膜H~+-ATPase活力的影响

采用两相法得到高纯度封闭的大豆下胚轴质膜微囊,研究了跨膜Ｃａ２＋梯差对质膜Ｈ＋－ＡＴＰａｓｅ质子转运和ＡＴＰ水解活力的影响。结果表明,在１０００：０．１,１０００：０．５,１０００：１及１０００：１０（μｍｏｌ／Ｌ：μｍｏｌ／Ｌ）几种梯差下,随着跨膜钙梯差的减小,质膜Ｈ＋－ＡＴＰａｓｅ质子转运活力逐步降低。然而,上述几种梯差对Ｈ＋－ＡＴＰａｓｅ水解活力的影响却很小。进一步研究发现,１０００：０．１及１０００：１（μｍｏｌ／Ｌ：μｍｏｌ／Ｌ）两种梯差对Ｋｍ值没有影响,但Ｋ＋对Ｈ＋－ＡＴＰａｓｅ的激活作用在两种梯差下存在显著差别。ＭＣ５４０荧光、ＤＰＨ荧光偏振结果表明,跨膜钙梯差影响着膜脂的聚集状态和流动性。本文对跨膜Ｃａ２＋梯差对于大豆下胚轴质膜Ｈ＋－ＡＴＰａｓｅ水解与质子转运活力影响的可能机制进行了讨论。     

温带阔叶红松林生态系统潜热通量模拟——气孔导度组合模型在Shuttleworth-Wallace双源模型中的应用

叶片水平的气孔导度组合模型已被成功扩展到冠层水平,并被应用于冬小麦生态系统潜热通量的模拟研究,但该研究仅基于1a的数据,有必要研究模型在更长时间尺度和其它生态系统类型的适用性.以长白山阔叶红松林(CBS)为研究对象,将组合模型进一步应用于Shuttleworth-Wallace双源模型,模拟了CBS 3a生长季内的潜热通量,利用涡度相关系统观测的潜热通量数据对模型进行验证,并对比了双源模型与单源模型的模拟结果.结果显示,双源模型较单源模型能取得更高的模拟精度,生长季不同时期的潜热通量模拟值和实测值的日变化较一致.对双源模型模拟值和实测潜热通量的相关分析显示,二者直线回归斜率和R2分别为0.96和0.72.对长白山阔叶红松林生态系统的蒸散和植被蒸腾的季节和年际变异分析发现,影响冠层蒸散和植被蒸腾季节动态的主要因素是饱和差和辐射,而影响它们年际动态的主要因素则是饱和差和温度.     

大鼠皂素蜕膜心肌肌钙蛋白Ca^2＋敏感性的实验研究

采用５００μｇ／ｍｌ皂素溶液浸浴大鼠右室乳头肌３０ｍｉｎ制备蜕膜心肌纤维标本。经撤除ＭｇＡＴＰ法检验,肌膜通透性明显增高。用含不同浓度Ｃａ￣(２＋)（以Ｃａ￣(２＋)浓度负对数ｐＣａ表示）的激活液进行顺序激活,记录Ｃａ￣(２＋)激活张力,其张力－ｐＣａ关系曲线描述了肌钙蛋白（ＴＮ）对Ｃａ￣(２＋)的亲和特性;曲线中位值ｐＣａ＿(５０)（即产生５０％最大张力所对应的ｐＣａ）是反映ＴＮＣａ￣(２＋)敏感性的定量指标。本实验观察到大鼠右室乳头肌张力－ｐＣａ关系曲线呈Ｓ形,测得ｐＣａ＿(５０)为５．７２７±０．０８６（ｎ＝１６）;撤除激活液中磷酸肌酸和磷酸肌酸激酶,张力－ｐＣａ曲线左移位,ｐＣａ＿(５０)增高至６．１９４±０．１２１（Ｐ＜０．０１）。     

体外培养新生大鼠大脑皮层星形神经元钾通道的特性及膜外钾离子浓度对其影响

用膜片钳技术中的细胞贴附方式和内面向外方式,首次在新生大鼠大脑皮层星形神经元胞体膜上记录到一类电压依赖性钾通道。此通道可被２０ｍｍｏｌ／ＬＴＥＡ,５ｍｍｏｌ／ＬＢａ２＋,１４０ｍｍｏｌ／ＬＣｓ＋阻断,不受２０ｍｍｏｌ／Ｌ４—ＡＰ影响,其激活不依赖Ｃａ２＋。膜外钾离子浓度对通道的特性有显著的影响,逆转电位随［Ｋ＋］０的增大而增大,并表现出一定的饱和现象,两者的对数呈线性关系;同一驱动电位下,平均开放时间和开放概率随［Ｋ＋］０的增大而增大,平均关闭时间的变化则相反。     

嗜菌紫膜表面电位和质子泵效率间的关系

本文研究了中性红与紫膜结合,并用直接滴定法和测定结合量法,求得了结合于膜上之中性红本征PK和在不同离子强度时的表观PKa值.从它们的差值中计算得到紫膜表面电位.并得不同盐浓度的表面电位和质子泵效率作了比较.结果说明.阳离子对质子泵效率的影响不能完全用表面电位的变化来说明.这一结果暗示了一定的阳离子对于紫膜质子泵功能的完成有着特殊作用.     

血红蛋白对人红细胞膜脂质分子侧向运动的影响

用荧光漂白恢复法测定了血红蛋白对红细胞膜脂质分子侧向扩散的限制作用.血红蛋白主要是通过和内侧膜脂质的结合而产生影响的,pH6及PH7.7时都显示出效应的存在.和膜结合较强的高铁血红蛋白,表现出对膜脂质侧向扩散亦有较大的限制作用.     

MICROTUBULATION OF THE INNER MEMBRANE OF THE NUCLEAR ENVELOPE

In the course of a light and electron microscopy study of spermatogenesis in the European crayfish, Astacus fluviatilis, spermatocytes of abnormal appearance were observed in two instances in individuals that had passed the mating period. The electron microscope showed that the inner membrane of the nuclear envelope of these cells was erupting into a mass of microtubules, 15 to 18 mµ in diameter and 0.5 µ or more in length, while the outer membrane transformed into cytoplasmic vesicles. Stages in the formation of these novel processes were followed. The plasma membrane of the affected cells was seen in some cases to erupt into similar although shorter microtubules. It is concluded that the phenomenon is part of a degenerative process in which the spermatocytes are being absorbed by sustentacular cells. It is suggested that the observations provide further evidence for a fundamental functional as well as a morphological similarity between the membranes bounding the nucleus and the plasma membrane.     

ON THE INFLUENCE OF AGGREGATES ON THE MEMBRANE POTENTIALS AND THE OSMOTIC PRESSURE OF PROTEIN SOLUTIONS

1. It is shown that when part of the gelatin in a solution of gelatin chloride is replaced by particles of powdered gelatin (without change of pH) the membrane potential of the solution is influenced comparatively little. 2. A measurement of the hydrogen ion concentration of the gelatin chloride solution and the outside aqueous solution with which the gelatin solution is in osmotic equilibrium, shows that the membrane potential can be calculated from this difference of hydrogen ion concentration with an accuracy of half a millivolt. This proves that the membrane potential is due to the establishment of a membrane equilibrium and that the powdered particles participate in this membrane equilibrium. 3. It is shown that a Donnan equilibrium is established between powdered particles of gelatin chloride and not too strong a solution of gelatin chloride. This is due to the fact that the powdered gelatin particles may be considered as a solid solution of gelatin with a higher concentration than that of the weak gelatin solution in which they are suspended. It follows from the theory of membrane equilibria that this difference in concentration of protein ions must give rise to potential differences between the solid particles and the weaker gelatin solution. 4. The writer had shown previously that when the gelatin in a solution of gelatin chloride is replaced by powdered gelatin (without a change in pH), the osmotic pressure of the solution is lowered the more the more dissolved gelatin is replaced by powdered gelatin. It is therefore obvious that the powdered particles of gelatin do not participate in the osmotic pressure of the solution in spite of the fact that they participate in the establishment of the Donnan equilibrium and in the membrane potentials. 5. This paradoxical phenomenon finds its explanation in the fact that as a consequence of the participation of each particle in the Donnan equilibrium, a special osmotic pressure is set up in each individual particle of powdered gelatin which leads to a swelling of that particle, and this osmotic pressure is measured by the increase in the cohesion pressure of the powdered particles required to balance the osmotic pressure inside each particle. 6. In a mixture of protein in solution and powdered protein (or protein micellæ) we have therefore two kinds of osmotic pressure, the hydrostatic pressure of the protein which is in true solution, and the cohesion pressure of the aggregates. Since only the former is noticeable in the hydrostatic pressure which serves as a measure of the osmotic pressure of a solution, it is clear why the osmotic pressure of a protein solution must be diminished when part of the protein in true solution is replaced by aggregates.     

用ANS研究脱落酸提高植物线粒体膜的流动性

用荧光标记法,以ＡＮＳ作荧光探剂研究了脱落酸（ＡＢＡ）对植被线粒体脱流动性的影响。（＋）ＡＢＡ浓度效应实验结果和不同ＰＨ及不同温度条件下的实验结果均表明（＋）ＡＢＡ能降低植物线粒体膜的荧光强度,即（＋）ＡＢＡ能提高线粒体原流动性,且此种效应具有浓度饱和性,并导致线粒体异柠檬酸脱氢酶（ＩＣＨ）活性升高。ＡＢＡ的类似物,ＲＣＡ．７Ａ和ＲＣＡ．７ｂ,也具有提高ＩＣＤＨ活性的效应,抗ＡＢＡ结合蛋白的抗体（     

Characteristics of tetraethylammonium transport in rabbit renal plasma-membrane vesicles.

Transport of [14C]tetraethylammonium (TEA), an organic cation, was studied in brush-border (BBMV) and basolateral (BLMV) membrane vesicles isolated from rabbit kidney cortex. In BBMV, the presence of an outwardly directed H+ gradient induced a marked stimulation of TEA uptake against its concentration gradient (overshoot phenomenon), whereas a valinomycin-induced inside-negative potential had no effect on TEA uptake. In BLMV, TEA uptake was significantly stimulated by the presence of an outwardly directed H+ gradient and by an inside-negative potential, but the effect of H+ gradient was absent when the vesicles were chemically 'voltage clamped'. In BBMV, internal H+ stimulated TEA uptake in a non-competitive manner by binding at a site with apparent pKa of 6.87. External H+ inhibited TEA uptake through a direct interaction with the putative H+/organic-cation exchanger at a site with apparent pKa of 6.78. Changing external pH while maintaining the pH gradient constant produced a result similar to that obtained by changing external pH alone. Increasing external H+ showed a mixed-type inhibition of TEA uptake. These results suggest that in the rabbit TEA transport across the basolateral membranes is driven by an inside-negative potential and that transport across the brush-border membrane is driven by a H+ gradient via an electroneutral H+/TEA antiport system.     

THE FIBRINOLYTIC ENZYME DEFECT OF HYALINE MEMBRANE DISEASE

An investigation of the pulmonary fibrinolytic enzyme system in 31 infants who died with hyaline membrane formation was reviewed. There was complete lack of plasminogen activator activity in the lungs of 84 per cent of these infants. This phenomenon was shown to result from an abnormal inhibitor. A comparable inhibitor was found in normal placental tissue, and it is postulated that this inhibitor is released into the circulating blood as the result of placental infarction. Fibrin, a basic component of the hyaline membrane, is probably precipitated from a physiological capillary transudate associated with the formation of amniotic fluid by the lungs. The presence of an inhibitor of fibrinolysis would then result in the accumulation of intrapulmonary fibrin and the formation of hyaline membranes.     

酰化对紫膜结构的影响

在很宽波长和ＰＨ范围内用紫外及可见区光吸收、圆二色及共振拉曼测定了酰化诱导的紫膜溶液的光谱变化。结构表明：酰化引起的表面电位的改变诱导了蛋白局部构象的改变。这种改变在很大程度上离域到整个蛋白分子中而影响了视黄醛结合位点的性质及生色团的结构,从而导致可观察的光谱变化。与未酰化紫膜比较,紫膜对ＰＨ的稳定性减小了。