ATP合酶:自然界最小的旋转发动机

ＡＴＰ合酶广泛存在于线粒体、叶绿体、原核藻、异养菌和光合细菌中,是生物体能量代谢的关键酶。该酶分别位于类囊体膜、质膜或线粒体内膜上,参与氧化磷酸化与光合磷酸化反应,在跨膜质子动力势的推动下催化合成生物体的能量“通货”——ＡＴＰ。ＡＴＰ合酶的Ｆ０部分比...     

Ⅲ°压疮皮肤组织iNOS、P53和细胞凋亡的变化

目的：观察Ⅲ°压疮创面组织中诱导型一氧化氮合酶（iNOS）与P53蛋白的表达以及细胞凋亡的情况,探讨Ⅲ°压疮慢性病程形成的原因。方法：在患者知情同意的情况下取9例患有Ⅲ°压疮病人的压疮溃疡组织,按压疮溃疡中心基底部组织、溃疡边缘组织进行分组,同时取4例手术遗弃的正常皮肤组织。采用免疫组织化学染色（SP法）,观察各组标本iNOS、P53的表达情况,采用末端标记技术（TUNEL）检测标本中的凋亡细胞。采用Image-prol-plus6.0计算机图像分析软件以积分光密度值（IOD）半定量检测iNOS、P53的表达强弱及凋亡细胞数目的多少。结果：压疮溃疡组织中心和边缘iNOS、P53的表达均强于正常皮肤组织,溃疡边缘组织表达强于正常皮肤组织（P均〈0.01）,溃疡中心组织iNOS的表达强于溃疡边缘组织（P〈0.01）,溃疡中心组织P53的表达强于溃疡边缘组织（P〈0.05）。从正常皮肤组织至溃疡边缘到溃疡中心,凋亡细胞数目显著增加（P〈0.01）,发生凋亡的细胞大部分是与修复相关的炎性细胞、成纤维细胞等。iNOS和P53与压疮组织中的细胞凋亡呈正相关,不控制P53时iNOS与细胞凋亡相关系数为0.605,不控制iNOS时P53与细胞凋亡相关系数为0.457,P均〈0.01。结论：Ⅲ°压疮创面组织中iNOS高表达产生高浓度的NO可能上调P53的表达,两者对创面组织中修复细胞的大量凋亡可能有促进作用,影响着压疮创面的愈合。     

高等植物环式电子传递的生理作用

环式电子传递做为一种可供选择的电子传递途径之一,近几年被证实它对于许多高等植物的生长是必需的.环式电子传递通过促进跨类囊体膜质子梯度的建立一方面激发ATP合成酶合成ATP,另一方面加强了光系统Ⅱ处的热耗散,稳定了放氧复合体,从而保护光系统Ⅱ免受光抑制.同时,它还可以缓解光系统Ⅰ处电子受体的过度还原,减少超氧阴离子在光系统Ⅰ处的合成,防止光系统Ⅰ受到光抑制.本文简要地综述了环式电子传递的途径、其参与ATP合成的作用、对光系统Ⅱ和光系统Ⅰ光保护作用及其对环境胁迫的响应和调节,并对环式电子传递的研究提出了展望.     

乳腺癌HPV16感染对iNOS、P53蛋白表达的影响

目的研究人乳头状瘤病毒（HPV16）在乳腺癌中的表达及作用机制,检测HPV16、诱导一氧化氮合酶（iNOS）及P53蛋白在乳腺癌的表达及其间的相关性。方法应用免疫组化SP法,共检测了52例乳腺癌和16例乳腺良性瘤HPV16、iNOS和P53蛋白的表达。结果HPV16和P53蛋白在乳腺癌组的阳性表达率均显著高于乳腺良性瘤组。统计分析表明,iNOS和P53蛋白表达阳性率与HPV感染率密切相关（P〈0．05）。结论HPV16感染参与了乳腺癌的发生,其致癌机制可能是通过诱导iNOS表达增加,产生NO,诱导p53基因突变,从而导致乳腺癌的发生。     

白血病细胞系来源的P2X7受体的功能研究

P2X7受体是ATP门控的离子通道。从白血病细胞系J6-1细胞中扩增P2X7受体编码区全序列,克隆到pTARGET真核表达载体,经DNA序列分析后转染Ramos细胞,获得稳定表达细胞株;应用RT-PCR、Westernblot和流式细胞术检测P2X7受体在Ramos中的表达;荧光分光光度计检测P2X7受体介导的胞内钙离子浓度变化。结果显示,J6-1细胞来源的P2X7受体在第559位有一个A→G的有义突变,导致Asn187→Asp187,可在Ramos细胞中表达,在特异性激动剂BzATP作用下可引起胞内钙离子浓度的升高,但所需激动剂浓度高于常规浓度。     

真菌中的脂质不对称调节和P4型ATP酶

脂质组分在双分子膜中不对称分布是广泛存在于真核细胞中的现象,有助于膜系统的出芽和融合,从而促进胞吞胞吐、蛋白分选运输等生理过程。多种蛋白参与维持脂质不对称,其中P4型ATP酶是重要的脂质转运体。P4型ATP酶又称脂质翻转酶,主要介导氨基磷脂的转位。在酿酒酵母、新生隐球菌、白念珠菌和构巢曲霉中,多种P4型ATP酶亚基的缺失导致蛋白运输失常,对唑类药物的敏感性升高,细胞壁完整性受损,毒力减低,在巨噬细胞中生存能力减弱等表型,提示脂质翻转酶的重要性及其作为抗真菌靶点的潜力。     

西藏虎头兰和墨兰对强光的生理响应

兰属植物(Cymbidium)是重要的观赏花卉。该属大多数种类生长在适度荫蔽的环境中,但附生型的西藏虎头兰(C.tracyanum)却能在阳光直射的环境下良好生长。为了探究西藏虎头兰适应强光的生理机制,研究选取同属的地生型墨兰(C.sinense)与其进行对照实验,测定了这两种植物在不同光照强度下的叶绿素荧光参数和P700氧化还原态。结果表明,在强光下,西藏虎头兰光系统I和II活性的下降程度比墨兰小,而环式电子传递的激发程度更高。环式电子传递的激发有助于增强西藏虎头兰在强光下的光保护,而墨兰因环式电子激发程度较低,使其不能适应强光。由于附生环境更容易出现间歇性的直射光,西藏虎头兰的这种适应强光的能力可能有助于它更充分地利用附生环境中的光照资源。     

西藏虎头兰和墨兰对强光的生理响应?

兰属植物( Cymbidium)是重要的观赏花卉。该属大多数种类生长在适度荫蔽的环境中,但附生型的西藏虎头兰( C?tracyanum)却能在阳光直射的环境下良好生长。为了探究西藏虎头兰适应强光的生理机制,研究选取同属的地生型墨兰( C?sinense)与其进行对照实验,测定了这两种植物在不同光照强度下的叶绿素荧光参数和P700氧化还原态。结果表明,在强光下,西藏虎头兰光系统I和II活性的下降程度比墨兰小,而环式电子传递的激发程度更高。环式电子传递的激发有助于增强西藏虎头兰在强光下的光保护,而墨兰因环式电子激发程度较低,使其不能适应强光。由于附生环境更容易出现间歇性的直射光,西藏虎头兰的这种适应强光的能力可能有助于它更充分地利用附生环境中的光照资源。     

稻瘟菌附着胞形成和发育的研究进展

附着胞是稻瘟菌侵染寄主的关键结构,cAMP、MAPK和Ca2+等信号途径参与其形成和发育,同时受寄主表面识别蛋白基因、黑色素合成基因、甘油合成基因、细胞自噬基因以及SNARE蛋白等因子调控。从以上几个方面综述了稻瘟菌附着胞形成与发育的研究进展。     

季节性干旱对白皮乌口树(Tarenna depauperata Hutchins)水分状况、叶片光谱特征和荧光参数的影响

干热河谷稀树灌丛常绿植物能够忍受长达半年以上的季节性干旱胁迫,但对这些常绿植物响应干旱胁迫的生理生态机制研究很少。本研究以干热河谷稀树灌丛优势常绿植物白皮乌口树(Tarenna depauperata Hutchins)为研究对象,分别在雨季和干季测定其叶片的水势、压力-体积曲线、气体交换参数、叶片光谱特征以及叶绿素荧光和P700的光能分配。结果显示:受严重季节性干旱胁迫的影响,与雨季相比,干季的凌晨叶片水势(Ψpd)下降至-4.5 MPa,水分传导的叶比导率(KL)下降了49.5%,叶绿素反射指数(NDVI)下降了40.6%,花青素反射指数(ARI)上升至0.074(约为雨季的12.3倍),并且雨季和干季的叶片水势、水分传导效率、叶绿素含量和花青素含量均差异显著(P0.05)。与雨季相比,干旱导致光系统Ⅱ(PSⅡ)最大光化学量子效率(Fv/Fm)显著下降至0.72(P0.05),即PSⅡ发生光抑制,而光系统Ⅰ(PSⅠ)的活性(Pm)未发生明显变化;干季叶片的最大非光化学淬灭(NPQ)增加了31%,而激发的最大环式电子传递速率(CEF)下降了66%。表明长期干旱胁迫使CEF的激发受到强烈抑制,即光能捕获效率的降低和NPQ的增强促进了白皮乌口树在长期干旱胁迫下的光保护。     

Integrating the proton circuit into photosynthesis: progress and challenges

The formation of trans-thylakoid proton motive force (pmf) is coupled to light-driven electron transfer and both powers the synthesis of ATP and acts as a signal for initiating antenna regulation. This key intermediate has been difficult to study because of its ephemeral and variable qualities. This review covers recent efforts to probe pmf in vivo as well as efforts to address one of the key questions in photosynthesis: How does the photosynthetic machinery achieve sufficient flexibility to meet the energetic and regulatory needs of the plant in a varying environment? It is concluded that pmf plays a central role in these flexibility mechanisms.     

The kinetics of the flash-induced P515 response in relation to the H+-permeability of the membrane bound ATPase in spinach chloroplasts

The effect of dicyclohexylcarbodiimide (DCCD) on the kinetics of the flashinduced P515 response and on the activity of the ATPase was investigated in isolated spinach chloroplasts. It was found that after the addition of 5×10^–8 mol DCCD the rate of ATP hydrolysis induced by a period of 60 sec illumination was decreased to less than 5% of its original value. At this concentration, hardly any effect, if at all, could be detected on the kinetics of the flash-induced P515 response, neither in dark-adapted nor in light-activated chloroplasts. It was concluded that the presence of concentrations of DCCD, sufficiently high to affect the ATPase activity, does not affect the kinetics of the flash-induced P515 response. Since DCCD decreases the H⁺ permeability of the membrane-bound ATPase, it was concluded that this permeability coefficient for protons is not an important factor in the regulation of the flash-induced membrane potential and, therefore, does not affect the kinetics of the flash-induced P515 response.     

苜蓿国际发展态势分析

苜蓿是草食动物的优质饲草, 被誉为“牧草之王”。发展苜蓿产业对提升我国草食畜牧业具有重要意义。该研究采用定性调研与定量分析相结合的方法, 从创新链角度, 研究了全球苜蓿科技产出、代表性国家苜蓿产业格局和全球苜蓿市场贸易等状况及我国苜蓿产业存在的问题, 旨在为我国苜蓿产业发展提供参考。分析发现, 美国是全球最重要的苜蓿生产国, 在苜蓿基础研究、技术开发、品种培育和商业化种植等方面均具有很强的优势, 引领了全球苜蓿产业的发展。欧美等跨国企业掌控着全球苜蓿产业链的各个关键环节, 是苜蓿产品的主要出口市场, 而亚洲苜蓿产品消费缺口最大。近10年来, 我国在苜蓿科技领域表现活跃, 科技成果产出呈快速增长趋势, 但在成果数量和影响力方面与欧美国家差距明显, 且苜蓿育种进程缓慢, 优质苜蓿产品对外依存度仍然较高。综合来看, 我国应持续加大苜蓿的研发力度和科技投入, 推进苜蓿产业化发展, 提升苜蓿产品的自给率, 保障草食畜牧业健康、稳定发展。     

P700氧化还原动力学的测量方法及原理

P700氧化还原动力学技术可快速且无损地检测植物光系统I (PSI)的活性, 是光合研究领域中广泛使用的一种技术。该文系统归纳了P700氧化还原动力学的主要测量方法, 详细阐述其原理并探讨该技术的局限性, 旨在为深入研究光合作用机理提供技术支持。     

The long-term responses of the photosynthetic proton circuit to drought

Proton motive force (pmf) across thylakoid membranes is not only for harnessing solar energy for photosynthetic CO₂ fixation, but also for triggering feedback regulation of photosystem II antenna. The mechanisms for balancing these two roles of the proton circuit under the long-term environmental stress, such as prolonged drought, have been poorly understood. In this study, we report on the response of wild watermelon thylakoid 'proton circuit' to drought stress using both in vivo spectroscopy and molecular analyses of the representative photosynthetic components. Although drought stress led to enhanced proton flux via a ∼34% increase in cyclic electron flow around photosystem I (PS I), an observed ∼fivefold decrease in proton conductivity, g_H⁺, across thylakoid membranes suggested that decreased ATP synthase activity was the major factor for sustaining elevated q_E. Western blotting analyses revealed that ATP synthase content decreased significantly, suggesting that quantitative control of the complex plays a pivotal role in down-regulation of g_H⁺. The expression level of cytochrome b ₆ f complex – another key control point in photosynthesis – also declined, probably to prevent excess-reduction of PS I electron acceptors. We conclude that plant acclimation to long-term environmental stress involves global changes in the photosynthetic proton circuit, in which ATP synthase represents the key control point for regulating the relationship between electron transfer and pmf.     

Alteration of bacterial surface electrostatic potential and pH upon adhesion to a solid surface and impacts to cellular bioenergetics

In our previous study [Hong Y, Brown DG (2009) Appl Environ Microbiol 75(8):2346–2353], the adenosine triphosphate (ATP) level of adhered bacteria was observed to be 2–5 times higher than that of planktonic bacteria. Consequently, the proton motive force (Δp) of adhered bacteria was approximately 15% greater than that of planktonic bacteria. It was hypothesized that the cell surface pH changes upon adhesion due to the charge‐regulated nature of the bacterial cell surface and that this change in surface pH can propagate to the cytoplasmic membrane and alter Δp. In the current study, we developed and applied a charge regulation model to bacterial adhesion and demonstrated that the charge nature of the adhering surface can have a significant effect on the cell surface pH and ultimately the affect the ATP levels of adhered bacteria. The results indicated that the negatively charged glass surface can result in a two‐unit drop in cell surface pH, whereas adhesion to a positively charged amine surface can result in a two‐unit rise in pH. The working hypothesis indicates that the negatively charged surface should enhance Δp and increase cellular ATP, while the positively charged surface should decrease Δp and decrease ATP, and these results of the hypothesis are directly supported by prior experimental results with both negatively and positively charged surfaces. Overall, these results suggest that the nature of charge on the solid surface can have an impact on the proton motive force and cellular ATP levels. Biotechnol. Bioeng. 2010;105: 965–972. © 2009 Wiley Periodicals, Inc.     

Differential effects of ambient or diminished CO2 and O2 levels on thylakoid membrane structure in light‐stressed plants

Over‐reduction of the photosynthetic electron transport chain may severely damage the photosynthetic apparatus as well as other constituents of the chloroplast and the cell. Here, we exposed Arabidopsis leaves to saturating light either under normal atmospheric conditions or under CO₂‐ and O₂‐limiting conditions, which greatly increase excitation and electron pressures by draining terminal electron acceptors. The two treatments were found to have very different, often opposing, effects on the structure of the thylakoid membranes, including the width of the granal lumenal compartment. Modulation of the latter is proposed to be related to movements of ions across the thylakoid membrane, which alter the relative osmolarity of the lumen and stroma and affect the partitioning of the proton motive force into its electrical and osmotic components. The resulting changes in thylakoid organization and lumenal width should facilitate the repair of photodamaged photosystem II complexes in response to light stress under ambient conditions, but are expected to inhibit the repair cycle when the light stress occurs concurrently with CO₂ and O₂ depletion. Under the latter conditions, the changes in thylakoid structure are predicted to complement other processes that restrict the flow of electrons into the high‐potential chain, thus moderating the production of deleterious reactive oxygen species at photosystem I.     

Measurement of components of proton motive force and related parameters in steady,microaerobic conditions

An experimental system is described for the simultaneous measurement of components of the proton motive force and other energy-related activies in microorganisms under steady defined microaerobic conditions. Oxygen is supplied in a solution containing an O₂-carrier such as myoglobin or leghaemoglobin, to a stirred reaction chamber in which a suspension of the microorganism is required aboce a membrane filter whic is pervious to the carrier. The rate of O₂ consumption is regulated by the rate at which the solution is pumped through the chamber. The concentration of free O₂ in the chamber and the rate of its consumption are calculated from the pumping rate and the decline in the relative oxygenation of the carrier measured spectrophotometrically in the effluent solution. The uptake by the microorganisms of radioactively labelled probes for ΔpH and Δψ is calculated following their injection into the reaction chamber and monitoring of continuously collected fractions of effluent solution, after it has passed through the spectrophometer. An example of the use of the system is given.The use of this system is advocated for many microaerobic applications since most of the measurements can be made without perturbing the steady state until the final shape of the suspension is collected.     

Direct interaction between linear electron transfer chains and solute transport systems in bacteria

Abstract In studies on alanine and lactose transport in Rhodopseudomonas sphaeroides we have demonstrated that the rate of solute uptake in this phototrophic bacterium is regulated by the rate of lightinduced cyclic electron transfer.
In the present paper the interaction between linear electron transfer chains and solute transport systems was studied in Rhodopseudomonas sphaeroides and Escherichia coli .
The results demonstrate that the activities of alanine transport in Rps. sphaeroides and lactose and proline transport in E. coli are directly controlled by the electron transfer activity in the respiratory chain, under conditions that the proton-motive force remains constant.     

Tentoxin effects on variable fluorescence and P515 electrochromic absorbance changes in tentoxin-sensitive and -resistant plant species

The effects of the cyclic tetrapeptide phytotoxin, tentoxin, on variable chlorophyll fluorescence quenching and the P515 electrochromic absorbance change were examined in tentoxin-sensitive and -resistant species and an interspecies hybrid. Immediately after infiltration of leaf discs, up to 100 μM tentoxin had no effect on the O, I, D, or P portions of the fluoresence transients of either tentoxin-sensitive or -resistant plants. However, the quenching of fluorescence following maximal fluorescence was reduced by approximately fourfold in tentoxin-treated tissues of tentoxin-sensitive plants, but was unaffected in resistant plants. Tentoxin significantly increased the magnitude of the P515 electrochromic absorbance change in all tentoxin sensitive plants by an average of approximately twofold. However, it was unaffected by tentoxin in resistant species. These data suggest that there is a close correlation between interaction of tentoxin with CF₁ ATPase in vivo and the ability to cause chlorosis in developing chloroplasts.