铜对小麦根质膜H+-ATPase活性的影响

用两相法提取小麦根质膜微囊,研究了铜对质膜H -ATPase活性的影响.硫酸铜(CuSO4)抑制小麦根质膜H -ATPase的活性,其作用具有浓度依赖性.金属离子螯合剂乙二胺四乙酸二钠(EDTA-Na2,200μmol.L-1)可以完全消除CuSO4对H -ATPase活性的抑制效应.检测硫代巴比妥酸反应物质(TBARS)的相对含量,结果显示Cu2 的加入使质膜微囊产生了较多的TBARS,表明Cu2 作用导致细胞膜发生较强的氧化损害.实验结果表明,CuSO4可以减弱小麦根质膜H -ATPase的活性,此效应是由Cu2 引起的;此外,Cu2 导致质膜微囊产生较强的膜脂过氧化.     

盐胁迫下植物质膜H+-ATPase研究进展

质膜H -ATPase在植物细胞的跨膜物质转运、胞内pH值调节以及植物对环境胁迫的响应等诸多方面具有重要作用,是植物生命活动过程的主宰酶.盐胁迫是影响植物生长发育的主要环境因子,植物质膜H -ATPase活性的调节是植物适应盐环境、提高耐盐性的重要细胞机理.综述了植物质膜H -ATPase活性变化及其调节机理对盐胁迫响应的研究状况,对质膜H -ATPase活性调节机理研究中仍未解决的问题进行了展望.     

H+-ATPase in the plasma membrane of Arabidopsis pollen cells is involved in extracellular calmodulin-promoted pollen germination

-activated Ca2+ channel in the PM of A.thaliana pollen cells.     

植物细胞膜H+ -ATPase对必需矿质养分的适应性

植物细胞膜H -ATPase是初级转运蛋白,在各种离子和代谢产物的跨膜运输中起着重要的作用;介绍了植物细胞H -ATPase的结构、生化特性、生理功能与调控机制;重点综述了细胞膜H -ATPase的活性及其蛋白数量与基因表达对植物必需矿质养分的适应性。     

植物细胞膜H+-ATPase对环境因子和逆境胁迫的适应

植物细胞膜H -ATPase是初级转运蛋白,它在养分离子跨膜运输、胞内pH调节、细胞伸长生长、气孔开闭,以及植物适应所处的环境与发育过程等生命活动中起着重要的作用.综述了植物细胞膜H -ATPase在酶活性、基因表达水平以及蛋白水平等方面对不同环境因子及逆境胁迫的适应性;指出分离鉴定、信号传导H -ATPase等将是进一步的研究方向.     

Targeting of transmembrane and GPI-anchored forms of N-CAM to opposite domains of a polarized epithelial cell.

The calcium-independent neural cell adhesion molecule N-CAM is expressed transiently during development in many tissues, including epithelia. The three naturally occurring principal isoforms of N-CAM differ in the way in which they associate with the membrane and in their cytoplasmic domains. These isoforms are generated by developmentally regulated alternative splicing of a single gene: the large cytoplasmic domain (ld) form (relative molecular mass 180,000 (Mr 180K] is specific for post-mitotic neurons; the 120K small cytoplasmic domain (ssd) and 140K small surface domain (sd) forms also occur on other cell types. One function of the different isoforms could be to specify cellular localization; for example, glycosyl phosphatidyl inositol (GPI)-membrane anchoring acts as a targeting signal for expression on the apical surface of polarized epithelial cells. Neurons and epithelial cells may use similar mechanisms for polarizing their plasma membrane proteins. We have therefore investigated the targeting of GPI-anchored (ssd N-CAM, 120K) and transmembrane forms of N-CAM (sd N-CAM, 140K; ld N-CAM, 180K) by comparing the expression of each after transfection of the appropriate complementary DNAs into polarized epithelial cells. We find that isoforms with alternative modes of membrane association are targeted to different surfaces of polarized epithelial cells: ssd N-CAM is expressed on the apical surface, whereas sd and ld N-CAM are expressed on the basolateral surface. These results suggest that the different isoforms of N-CAM determine their own diverse cellular destinations. They also support the hypothesis that the GPI anchor acts as an apical targeting signal in epithelia.     

钙对玉米幼苗的抗寒性及膜H+-ATPase活性的影响

用50mmol/L CaCl2浸泡玉米种子36h之后，将其幼苗放在低温下胁迫处理，蒸馏水处理作对照。结果表明：处理组玉米幼苗的成活率明显高于对照组，而且，处理组的质膜、液膜膜H^ -ATPase活性也明显高于对照组；但处理组电解质渗透率却低于对照组。说明Ca^2 能提高玉米幼苗的抗寒性，也能提高质膜、液泡膜H^ -ATPase活性，能降低叶片的电解质渗透率。     

植物细胞膜H -ATPase对必需矿质养分的适应性

植物细胞膜H -ATPase是初级转运蛋白,在各种离子和代谢产物的跨膜运输中起着重要的作用;介绍了植物细胞H -ATPase的结构、生化特性、生理功能与调控机制;重点综述了细胞膜H -ATPase的活性及其蛋白数量与基因表达对植物必需矿质养分的适应性.     

仙人掌多糖对荷瘤小鼠肿瘤细胞钙泵的影响

研究了食用仙人掌多糖、药用仙人掌多糖对S180小鼠肿瘤细胞膜Ca2 -ATPase的活性影响.结果表明,二者均能明显抑制荷瘤小鼠肿瘤细胞膜上Ca2 -ATPase的活性.仙人掌多糖的这一作用改变了荷瘤小鼠细胞膜的物质、能量平衡,推测其作用可能表现为促进肿瘤细胞的凋亡.     

Ca2+-activated ATPase and ATP-dependent calmodulin-stimulated Ca2+ transport in islet cell plasma membrane

Calcium is known to play an essential part in the regulation of insulin secretion in the pancreatic beta cell. Calcium influx/efflux studies indicate that glucose promotes an accumulation of calcium by the beta cell. However, interpretation of such data is particularly difficult due to the complex compartmentalization of calcium within the cell. Although indirect evidence using chlorotetracycline suggests that control of calcium homeostasis at the plasma membrane may be central to insulin secretion, the mechanism by which secretagogues influence the handling of calcium remains unknown. Despite its continuous diffusive entry, intracellular calcium is maintained in the submicromolar range by energy-dependent mechanisms. One such process which has been well characterized in erythrocytes is a plasma membrane calcium extrusion pump whose enzymatic basis is a high affinity (Ca+2 + Mg+2)ATPase. A similar mechanism regulated by insulin has recently been identified in adipocyte plasma membranes. We report here the presence of a high affinity (Ca+2 + Mg+2)ATPase and ATP-dependent calmodulin-stimulated calcium transport system in rat pancreatic islet cell plasma membranes.     

Evidence for polarization of plasma membrane domains in pancreatic endocrine cells

Polarization of plasma membrane domains is an essential feature of secretory epithelial cells from exocrine glands. The surface of exocrine cells (a typical example is the acinar cell of the pancreas) is separated into an apical domain, where secretion occurs by exocytosis, and a basolateral domain, which senses variations of the internal milieu and is enriched with receptors for various hormones and secretagogues. It is unknown whether secretion is polarized in endocrine cells (except for thyroid follicular cells, which are organized into cavitary structures). To determine whether distinct plasma membrane domains exist in endocrine cells, we infected monolayer cultures of pancreatic endocrine cells with enveloped RNA viruses known to bud selectively from either the apical or basolateral domain in polarized epithelial cells. This asymmetrical budding is thought to reflect the polarized nature of the infected cells, as in non-polarized cells such as fibroblasts, the same viruses bud nonselectively from the entire cell surface. We show here that influenza virus and vesicular stomatitis virus (VSV) emerge asymmetrically from cultured pancreatic islet cells; this represents the first evidence for polarization of plasma membrane domains in pancreatic endocrine cells.     

芦荟多糖对荷瘤小鼠肿瘤细胞钠泵的影响

报道了库拉索芦荟多糖、木立多糖对S1 80 小鼠肿瘤细胞膜Na+ ,K+ -ATPase的活性影响。结果表明 ,二者均能明显抑制荷瘤小鼠肿瘤细胞膜上Na+ ,K+ -ATPase的活性。芦荟多糖的这一作用改变了荷瘤小鼠细胞膜的物质、能量平衡 ,进而降低了肿瘤细胞的恶性程度。     

Carbon-dioxide-induced exocytotic insertion of H+ pumps in turtle-bladder luminal membrane: role of cell pH and calcium

The contents of endocytic vesicles and other intracellular organelles (such as Golgi and microsomes) are acidified by an electrogenic proton-translocating ATPase that is remarkably similar to that found in urinary epithelia. We recently found that the number of H+ ATPases in the apical plasma membrane of these epithelia is regulated by exocytotic insertion of endocytic vesicles whose membranes contain this H+ pump. Carbon dioxide, a major stimulus for urinary acidification, causes rapid fusion of these vesicles with the luminal membrane, thereby inserting these pumps there and increasing the rate of net transepithelial H+ secretion; CO2 also inhibits endocytic retrieval of the pumps from the luminal membrane. Such reciprocal regulation of endocytosis and exocytosis by a physiological modulator makes this system particularly attractive for studying the cellular events regulating membrane fusion. Here we present evidence that CO2 induces exocytosis by a cascade of events, the first step of which is cytoplasmic acidification. Cell acidification then increases calcium activity, which causes the fusion event.     

Epstein-Barr virus latent membrane protein inhibits human epithelial cell differentiation

Epstein-Barr virus (EBV), a human herpesvirus, is strongly linked with two relatively rare forms of B-cell lymphoma and with a much more prevalent epithelial malignancy, undifferentiated nasopharyngeal carcinoma (NPC). The availability of suitable culture systems has allowed detailed analysis of EBV-induced growth transformation in B lymphocytes, but little is known about the virus--epithelial cell interaction or about the possible effector role of viral proteins in the pathogenesis of NPC. Here we describe an experimental system to monitor the effects of introduced viral or cellular genes upon human epithelial cell growth and differentiation. We transfected a human epithelial cell line, which retains several features of normal keratinocyte behaviour in vitro, with the EBV gene encoding latent membrane protein (LMP), one of only two viral proteins known to be expressed in NPC cells in vivo. LMP expression was accompanied by changes in the epithelial cell surface phenotype, mimicking surface changes observed in NPC cells, and by severe impairment of the cellular response to differentiation signals. The ability of LMP to inhibit terminal differentiation indicates a mechanism whereby EBV infection of squamous epithelium could contribute to the multi-step pathogenesis of NPC.     

胰蛋白酶处理对大豆下胚轴质膜H+-ATPase的影响

以大豆下胚轴为材料,采用蔗糖密度梯度法制备高纯度质膜微囊,研究了胰蛋白酶处理对质膜Ｈ＾＋－ＡＴＰａｓｅ的影响。     

穿山龙液泡H+-ATPase c亚基基因片段的克隆

利用抑制消减杂交（SSH）和SMART技术，构建了3年生和1年生穿山龙（Dioscorea nipponica）根组织mRNA群体间正向差减cDNA文库．从34个随机测序的cDNA克隆中，发现了1个编码液泡H^＋-ATPase（V-H^＋-ATPase）c亚基的克隆，并命名为DnvHAc1（Accession No：DN792564）．序列同源性分析表明，其cDNA序列长486bp，3’端具有PolyA结构，其编码的氨基酸序列（115个氨基酸残基）也与多种植物的液泡H^＋-ATPase c亚基（16kDa proteolipids）具有较高同源性，具有3个跨膜疏水结构域，结构域Ⅲ为功能保守的区域，有dicyclohexylcarbodimide（DCCD）结合位点，Glu-92在调节液泡H^＋-ATPase具有重要作用．该研究结果为克隆其全长基因和进一步研究其在薯蓣皂甙次生代谢生物合成中的功能提供了重要的实验基础．