叶绿体类囊体膜蛋白质磷酸酯酶

概述了植物蛋白质磷酸酯酶的研究现状,重点介绍了叶绿体类囊体膜蛋白质磷酸酯酶的分离、纯化、定位、影响酶活力的因子,以及酸在类囊体膜蛋白质磷酸化调节中表现出的多样性。     

冷害对黄瓜叶绿体类囊体膜的影响

研究了冷害温度(0℃,16h)对黄瓜(Cucumis sativus L.)叶绿体类囊体膜膜脂、膜蛋白成分的影响。在没有可见伤害症状的低温处理条件下,黄瓜叶片叶绿体类囊体膜膜脂成分已有变化,主要是磷脂酰甘油(PG)含量明显降低,但主要脂类成分单半乳糖基甘油二酯(MGDG)、双半乳糖基甘油二酯(DGDG)、硫代异鼠李糖甘油二酯(SQDC)和PG的脂肪酸组分没有明显的变化;类囊体膜上色素蛋白质复合体的变化以光系统Ⅱ捕光叶绿素a/b蛋白质(LHCⅡ)单体及寡聚体含量的变化最明显,低温处理使LHCⅡ单体比例增加。对提纯的LHCⅡ结合脂的分析表明,低温处理改变了LHCⅡ结合脂及其脂肪酸的组成,使PG含量降低。以上结果表明,LHCⅡ结合脂成分变化以及LHCⅡ寡聚体解聚可能是叶绿体类囊体膜受冷害的最初反应。     

植物叶绿体类囊体膜及膜蛋白研究进展

叶绿体是植物和真核藻类进行光合作用的场所。存在于叶绿体类囊体膜上的蛋白质复合物含有光反应所需的光合色素和电子传递链组分,在光合作用过程中,光化学反应发生在类囊体膜上。因此,类囊体膜是光能向化学能转化的主要场所,因而也一直是光合作用研究的热点。叶绿体类囊体膜的深入研究可以促进光合作用的分子机理研究。该文就叶绿体类囊体膜的三维构象及类囊体膜蛋白的组成和功能研究进行了综述。     

叶绿体类囊体膜蛋白质的化学交联效应

研究了蛋白质化学交联剂二氟二硝基苯对叶绿体类囊体膜功能的效应。检测到ＤＦＤＮＢ抑制光合磷酸化,降低光照诱导类囊体膜的质子吸收和９－氨基吖啶（９－ＡＡ）的荧光猝灭,降低电色效应的快相上升,显抑制膜上腺三磷酶的活性,ＤＦＤＮＢ和游离的腺三磷酶进行交联反应,再经ＳＤＳ－聚丙烯酰胺凝胶电泳表明,交联后的酶出现新的蛋白染色带。     

热处理对菠菜和上海青类囊体膜及叶绿素稳定性的影响

绿色蔬菜在加工和贮藏过程中发生的颜色劣变与类囊体膜上叶绿素蛋白复合体的稳定性有着密切的关系。本研究以上海青和菠菜为原料,研究了两种绿色蔬菜在热处理过程中类囊体膜溶液热容、多肽组分、叶绿素含量、pH以及蛋白质荧光的变化。结果表明：热处理会导致叶绿素蛋白复合体的降解,类囊体膜溶液pH下降,上海青类囊体膜热稳定性、叶绿素含量的保持优于菠菜类囊体膜。试验结果为我们明确绿色蔬菜颜色劣变的机理以及寻求更好的护绿手段提供了理论基础。     

低温弱光对黄瓜和菠菜类囊体跨膜质子梯度的影响

以典型冷敏性植物黄瓜和典型抗冷性植物菠菜为材料 ,研究了 5℃ 1 0 0μmol pho-tons· m-2 · s-1 低温弱光处理对 2种植物的活体和离体叶绿体类囊体耦联度的影响。结果显示 ,与 5℃黑暗处理相比 ,5℃下弱光 (1 0 0 μmol photons·m-2· s-1 )分别照射黄瓜和菠菜的叶片和离体叶绿体悬浮液 ,都使叶绿体毫秒延迟发光慢相强度以及类囊体耦联度显著降低。表明无论是冷敏性作物黄瓜 ,还是抗冷性植物菠菜 ,低温弱光处理叶片和离体叶绿体悬浮液 ,均可导致类囊体跨膜质子梯度显著降低     

CO2浓度增加对玉米杂交种和亲本叶绿体类囊体膜光合特性的影响

当今世界工农业生产迅速发展,人口急剧增加,对能源的消耗也与日俱增。因此大气层中的 CO_2浓度不断升高。有专家预计,到21世纪后期,全球的 CO_2浓度将会升高一倍。伴随着 CO_2浓度的升高,全球的气温也将升高5—6℃。随着大气和温度的变化,其他气候因子也会发生相应变化,这些都将给植物生长发育带来预想不到的影响。因此 CO_2浓度     

Isolation and Characterization of Phosphatase from Thylakoid Membranes of Spinach Chloroplasts

A phosphatase from thylakoid membrane of spinach (Spinacia oleracea L. ) chloroplasts was isolated with the methods of extraction with n-ButanoL centrifugation at 100000 g for 30 min and chromatographic separation through DEAE-Cellulose (DE 52) column.The phosphatase catalyzed hydrolysis of phosphate monoesters (4-nitrophenyl phosphate). The optimal pH for enzyme catalysis was below 7. The peak rate of the enzyme reaction was obtained when it was incubated at 60℃ for 15 min. The phosphatase was inhibited by ATP and phosphate. The results from SDS-PAGE showed that the preparation of enzyme was composed of two proteins.     

野生和黄化大麦类囊体膜色素蛋白的分离和比较

用混和去垢剂（ＳＤＳ、ＯＧ）增溶和温和电泳方法,对野生和黄化大麦的叶绿体类囊体膜进行了色素蛋白复合体组成的分离和比较,并结合ＳＤＳ－聚丙烯酰胺凝胶电泳对类囊体膜蛋白进行了鉴定。结果表明,黄化大麦类囊体膜色素蛋白组分中,ＬＨＣ Ⅱ和ＬＨＣ Ⅰ的含量都较野生型大麦明显减少;并讨论了黄化大麦类囊体膜色素蛋白含量的减少与其光合速率降低的关系。     

ISOLATION AND CHARACTERIZATION OF LUMINAL MEMBRANES FROM URINARY BLADDER

A method is reported for the isolation of a highly purified fraction of urinary bladder membranes containing hexagonal plaques. The method uses zonal centrifugation as the final step of fractionation. The purified fraction was characterized by its electron microscopic morphology, by its enzymatic profile, by quantitative and qualitative analysis of lipids and by the protein pattern obtained by electrophoresis in polyacrylamide sodium dodecyl sulfate gels. The fraction contains 65% lipids and 35% proteins. The major protein component has a molecular weight of 27,000 daltons. Phospholipids are more than the 54% of the total lipid weight. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol are the major phospholipids with 50%, 30%, and 7% of the total lipid phosphorus, respectively. The glycolipid fraction is 10% of the total lipid weight and is formed by only two components, both sulfatides. Total cholesterol makes up 36% of the total neutral lipid fraction of which cholesterol esters constitute 6%. Glycoproteins are also found to be present in the fraction.     

菠菜叶片类囊体膜上K^＋通道的直接测定及其离子选择性

将经蔗糖密度梯度离心纯化的菠菜（Ｓｐｉｎａｃｅａ ｏｌｅｒａｃｅａ Ｌ．） 叶片类囊体膜微囊泡嵌入平面脂双层（ｐｌａｎａｒ ｌｉｐｉｄ ｂｉｌａｙｅｒｓ）,至少检测到一种Ｋ＋ 单通道信号, 其电导约为５５ ｐＳ,具有电压依赖特性,对ＴＥＡ（ｔｅｔｒａｅｔｈｙｌａｍ ｍ ｏｎｉｕｍ ｃｈｌｏｒｉｄｅ）敏感,这一通道的ＰＫ＋ ／ＰＣｌ－ 通透比约为１４．９。离子通道的选择性可以在对称的离子条件下测定不同离子的电导,或者在双离子条件下测定反转电位确定,其对单价阳离子的选择顺序为：Ｋ＋ ＞ Ｎａ＋ ＞ Ｌｉ＋ ＞ ＮＨ＋４ ＞ Ｃｓ＋ 。     

ISOLATION AND CHARACTERIZATION OF MYELIN-RELATED MEMBRANES

Abstract— Myelin related membrane fractions from rat brain and spinal cord were isolated from material normally discarded during standard myelin isolation procedures. A fraction which floated on 0.32 M-sucrose (F) and the material released after subjecting the myelin fraction to osmotic shock at two stages in the purification (W₁ and W₂) were characterized. These fractions were subjected to subfractionation on three step discontinuous sucrose gradients. Morphologically, the heavier subfrac-tions of W₁ and W₂ were shown to consist mainly of single membranes and vesicles. Sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis showed that, relative to myelin, proteolipid and basic protein were reduced in all subfractions, while the high molecular weight proteins were increased. The specific activity of 2′,3′-cyclic nucleotide 3′-phosphohydrolase (CNP) was up to 2-fold higher than that of myelin in the heavier subfractions of W₁ and W₂. The major myelin-associated glycoprotein was also increased in these subfractions as determined by periodic acid-Schiff staining. Differential centrifugation of the initial tissue homogenate to remove microsomes prior to myelin isolation gave rise to W₁ and W₂ subfractions with a CNP specific activity 3–4 times that of myelin. The high molecular weight proteins and glycoproteins were enriched in these microsome-depleted subfractions, but were qualitatively similar to those of myelin. Some of the membranes in these fractions may be derived from the continuum between the plasma membrane of the oligodendrocyte and compact myelin. Fraction F consisted of small membrane fragments and many vesicles, and was particularly deficient in proteolipid. The specific activity of CNP in fraction F was about the same as myelin, while the major myelin associated glycoprotein could not be detected. Fraction F from normal CNS tissue appears to be similar to the floating fractions previously isolated in larger amounts from pathological brain undergoing edematous demyelination.     

光周期对光敏核不育水稻叶绿体类囊体膜超分子结构的影响

通过冰冻撕裂叶绿体类囊体膜电镜术的研究,在超分子水平上揭示出长日照（ＬＤ）与短日照（ＳＤ）对突变型光敏核雄性不育水稻农垦５８Ｓ及其野生型农垦５８ 倒二叶叶绿体类囊体膜的形成有不同的影响。农垦５８－ＳＤ 和农垦５８－ＬＤ差别不明显;而农垦５８Ｓ－ＳＤ和农垦５８Ｓ－ＬＤ之间出现较明显的差异：（１）农垦５８Ｓ－ＳＤ 叶绿体类囊体膜的垛叠和非垛叠膜区的４ 个冰冻撕裂膜小面呈现的功能蛋白粒的密度、大小及构象分布与其对照农垦５８－ＳＤ 和农垦５８－ＬＤ 类似,均属正常的超分子结构类型。（２）农垦５８Ｓ－ＳＤ叶绿体垛叠类囊体膜区的外质撕裂面（ＥＦｓ）功能蛋白粒的密度比５８Ｓ－ＬＤ的大,且在有的垛叠膜区呈现出类晶格状的规则排列。（３）农垦５８Ｓ－ＬＤ 叶绿体非垛叠膜区的原生质撕裂面（ＰＦｕ）和外质撕裂面（ＥＦｕ）出现频率较低,往往在基粒的边缘膜或末端中能找到,且分布于膜上的功能蛋白粒的密度较小。有的叶绿体ＥＦｕ、ＰＦｕ、ＥＦｓ和垛叠膜区原生质撕裂面（ＰＦｓ）功能蛋白粒极少或丧失,表明类囊体膜受到严重损伤     

镉离子对菠菜叶绿体色素蛋白质复合物及激发能分配的影响

Cd~(2+)使叶绿体低温(77K)荧光发射光谱中 F686/F736及 F696/F736和激发光谱中F480/F436比值降低,说明 Cd~(2+)不利于激发能向 PSⅡ传递。SDS-PAGE 分析表明,Cd(2+)处理后叶绿体类囊体膜中光系统Ⅱ捕光叶绿素蛋白质复合物 LHCⅡ的部分寡聚体解聚成单体,且 LHCⅡ的总量也减少了。分析表明 Cd~(2+)使属于 LHCⅡ的多肽减少。已知LHCⅡ在光能吸收、传递以及激发能在两个光系统间的分配和调节方面起着重要作用,Cd~(2+)引起部分 LHCⅡ解聚和总量减少,必然导致由 LHCⅡ捕获和向光系统Ⅱ中心传递的能量减少。     

玉米类囊体膜的超分子结构及LHCP在个体发育中的变化

对玉米(Zea mays)营养生长期中的下位叶(第5叶)和生殖生长时期的中位叶(果穗叶)和上位叶(顶生叶)的成熟叶片的冰冻撕裂电镜观察,发现叶绿体类囊体膜所有撕裂面上各种功能蛋白颗粒的密度均以果穗叶中的最大,依次是顶生叶和第5叶的。以果穗叶与顶生叶相比,其类囊体膜中包含有绝大多数 LHCP 的 EFs 颗粒增加28%;包含有 PSI 反应中心与LHCP 相结合的 PFu 颗粒增加20%;包含有 PSII 反应中心与 LHCP 相结合的 EFs 颗粒增加19%。这一超分子结构的电镜观察结果与其 SDS-聚丙烯酰胺梯度凝胶板电泳解析的结果相一致。即 SDS-聚丙烯酰胺梯度凝胶板电泳解析的色素带上,同样是果穗叶类囊体膜上呈现的21kD(LHCP Ⅰ)和25 kD(LHCPⅡ)多肽的色素带相应地也比顶生叶的加宽,表明果穗叶叶绿体类囊体膜上镶嵌的叶绿素 a/b 蛋白复合体等比顶生叶的显著地增多,这有利于果穗叶光合作用中光能的吸收、传递、分配和转化。     

盐胁迫对大麦叶片类囊体膜上两种形态多胺含量和膜蛋白合成的影响

 用不同浓度NaCl处理6天龄大麦（Hordeum vulgare）幼苗3 d,以非共价键形式与叶片类囊体膜结合的多胺含量在低浓度盐处理时含量不变,随NaCl处理浓度的上升含量明显下降。在检测到的3种多胺中Spd（亚精胺）含量最高,约占膜上非共价键结合多胺总量的40%。以共价键形式与类囊体膜蛋白结合的多胺含量随NaCl处理浓度的上升一直呈下降趋势。在检测到的3种多胺中Put（腐胺）含量最高,约占膜上共价键结合多胺总量的60%以上。低浓度盐处理对14C-Glu向类囊体膜蛋白的掺入没有影响,随NaCl处理浓度的上升14C-Glu向类囊体膜蛋白的掺入明显下降。