Molecular mechanism of vectorial proton translocation by bacteriorhodopsin

Bacteriorhodopsin, a membrane protein with a relative molecular mass of 27,000, is a light driven pump which transports protons across the cell membrane of the halophilic organism Halobacterium salinarum. The chromophore retinal is covalently attached to the protein via a protonated Schiff base. Upon illumination, retinal is isomerized. The Schiff base then releases a proton to the extracellular medium, and is subsequently reprotonated from the cytoplasm. An atomic model for bacteriorhodopsin was first determined by Henderson et al, and has been confirmed and extended by work in a number of laboratories in the last few years. Here we present an atomic model for structural changes involved in the vectorial, light-driven transport of protons by bacteriorhodopsin. A 'switch' mechanism ensures the vectorial nature of pumping. First, retinal unbends, triggered by loss of the Schiff base proton, and second, a protein conformational change occurs. This conformational change, which we have determined by electron crystallography at atomic (3.2 A in-plane and 3.6 A vertical) resolution, is largely localized to helices F and G, and provides an 'opening' of the protein to protons on the cytoplasmic side of the membrane.     

冶金反应过程动力学的分段尝试法

冶金反应工程学属于一门独立的学科,需要建立自己的一套研究反应动力学的新方法。为了避免与冶金物理化学的方法和内容的重叠,本文提出分段尝试法可作为研究冶金反应过程动力学的新方法。依据反应过程特点,建立相应动力学模型进行拟合尝试,能够得到该过程控制环节转换时间点以及动力学参数,为冶金模拟提供必要的物性参数。由于在实际冶金反应过程中,化学反应过程和质量传输过程的耦合是客观存在的,化学反应控制时质量传输也发生;同理,质量传输控制时也有化学反应。因此,提出通过引用不可逆过程热力学唯象理论来确定出化学反应和扩散控制的两种通量之间的影响,从而进一步完善分段尝试法。     

Effects of the tumour promoter okadaic acid on intracellular protein phosphorylation and metabolism

Okadaic acid is a polyether derivative of 38-carbon fatty acid, and is implicated as the causative agent of diarrhetic shellfish poisoning. It is a potent tumour promoter that is not an activator of protein kinase C, but is a powerful inhibitor of protein phosphatases-1 and -2A (PP1 and PP2A) in vitro. We report here that okadaic acid rapidly stimulates protein phosphorylation in intact cells, and behaves like a specific protein phosphatase inhibitor in a variety of metabolic processes. Our results indicate that PP1 and PP2A are the dominant protein phosphatases acting on a wide range of phosphoproteins in vivo. We also find that okadaic acid mimics the effect of insulin on glucose transport in adipocytes, which suggests that this process is stimulated by a serine/threonine phosphorylation event.     

Plasticity of functional epithelial polarity

The fundamental characteristics that allow vectorial transport across an epithelial cell are the differential sorting and insertion of transport proteins either in the apical or the basolateral plasma membrane, and the preferential association of endocytosis and exocytosis with one or the other pole of the cell. Asymmetrical cellular structure and function, being manifestations of terminal differentiation, might be expected to be predetermined and invariant. Here we show that the polarity of transepithelial H+ transport, endocytosis and exocytosis in kidney can be reversed by environmental stimuli. The HCO3- secreting cell in the cortical collecting tubule is found to be an intercalated cell possessing a Cl-/HCO3- exchanger in the apical membrane and proton pumps in endocytic vesicles that fuse with the basolateral membrane; the H+-secreting cell in the medullary collecting tubule has these transport functions on the opposite membranes. Further, the HCO3- -secreting cell can be induced to change its functional polarity to that of the H+-secreting cell by acid-loading the animal.     

复合气液传质过程多元传质系数估算方法

以多元传质的扩散方程为基础，提出化学反应，吸附对气液传质增强因子以及伴有化学反应、吸附的气液传质过程的多元传质系数的估算方法，用非平衡级模型及上述方法得到的传质系模拟脂肪酸甲酯化反应精馏过程，与实验结果吻合。     

Role of CO2 in the formation of gold deposits

Much of global gold production has come from deposits with uneconomic concentrations of base metals, such as copper, lead and zinc. These 'gold-only' deposits are thought to have formed from hot, aqueous fluids rich in carbon dioxide, but only minor significance has been attached to the role of the CO2 in the process of gold transport. This is because chemical bonding between gold ions and CO2 species is not strong, and so it is unlikely that CO2 has a direct role in gold transport. An alternative indirect role for CO2 as a weak acid that buffers pH has also appeared unlikely, because previously inferred pH values for such gold-bearing fluids are variable. Here we show that such calculated pH values are unlikely to record conditions of gold transport, and propose that CO2 may play a critical role during gold transport by buffering the fluid in a pH range where elevated gold concentration can be maintained by complexation with reduced sulphur. Our conclusions, which are supported by geochemical modelling, may provide a platform for new gold exploration methods.     

Functional relationship of cytochrome c(6) and plastocyanin in Arabidopsis

Photosynthetic electron carriers are important in converting light energy into chemical energy in green plants. Although protein components in the electron transport chain are largely conserved among plants, algae and prokaryotes, there is thought to be a major difference concerning a soluble protein in the thylakoid lumen. In cyanobacteria and eukaryotic algae, both plastocyanin and cytochrome c(6) mediate electron transfer from cytochrome b(6)f complex to photosystem I. In contrast, only plastocyanin has been found to play the same role in higher plants. It is widely accepted that cytochrome c(6) has been evolutionarily eliminated from higher-plant chloroplasts. Here we report characterization of a cytochrome c(6)-like protein from Arabidopsis (referred to as Atc6). Atc6 is a functional cytochrome c localized in the thylakoid lumen. Electron transport reconstruction assay showed that Atc6 replaced plastocyanin in the photosynthetic electron transport process. Genetic analysis demonstrated that neither plastocyanin nor Atc6 was absolutely essential for Arabidopsis growth and development. However, plants lacking both plastocyanin and Atc6 did not survive.     

茶籽油和饼粕的化学成分研究

本文报道了茶籽的化学成分和利用价值.经测定,茶子中粗蛋白、粗脂肪和粗皂甙的含量分别为9.1%、21.9%和15.4%.茶籽油在理化特性和脂肪酸组成方面与油茶籽油十分接近.茶籽油的酸值、碘值和皂化值分别为1.82、89.41和191.22.茶籽油的脂肪酸有棕榈酸(C_(16∶0))、硬脂酸(C_(18∶0))、花生酸(C_(20∶0))、棕榈油酸(C_(16∶1))、油酸(C_(18∶1))、亚油酸(C_(18∶2))和亚麻酸(C_(18∶3))等七个组分.茶籽油中,不饱和脂肪酸为75.89%,其中油酸和亚油酸占73.83%,并测定了茶籽饼粕的化学成分和氨基酸组成.在其饼粕中,粗蛋白含量为16.4%,必需氨基酸齐全且含量较高。研究结果表明茶籽饼粕可以作为一种新的饲料资源加以利用.     

Mechanically driven ATP synthesis by F1-ATPase

ATP, the main biological energy currency, is synthesized from ADP and inorganic phosphate by ATP synthase in an energy-requiring reaction. The F1 portion of ATP synthase, also known as F1-ATPase, functions as a rotary molecular motor: in vitro its gamma-subunit rotates against the surrounding alpha3beta3 subunits, hydrolysing ATP in three separate catalytic sites on the beta-subunits. It is widely believed that reverse rotation of the gamma-subunit, driven by proton flow through the associated F(o) portion of ATP synthase, leads to ATP synthesis in biological systems. Here we present direct evidence for the chemical synthesis of ATP driven by mechanical energy. We attached a magnetic bead to the gamma-subunit of isolated F1 on a glass surface, and rotated the bead using electrical magnets. Rotation in the appropriate direction resulted in the appearance of ATP in the medium as detected by the luciferase-luciferin reaction. This shows that a vectorial force (torque) working at one particular point on a protein machine can influence a chemical reaction occurring in physically remote catalytic sites, driving the reaction far from equilibrium.     

Selective dendritic transport of RNA in hippocampal neurons in culture

Typical neurons of the central nervous system (CNS) elaborate tens of thousands of membrane specializations at sites of synaptic contacts on their dendrites. To construct, maintain, and modify these specializations, neurons must produce and deliver the appropriate molecular constituents to particular synaptic sites. Previous studies have revealed that polyribosomes are selectively positioned beneath postsynaptic sites, suggesting that in neurons, as in other cell types, protein synthetic machinery is located at or near the sites where particular proteins are needed. The mechanisms that deliver ribosomes and messenger RNA to their specific destinations in cells are therefore of considerable interest. Here we describe a system for RNA transport in dendrites that could provide a mechanism for the delivery of ribosomes and mRNA to synaptic sites in dendrites. Hippocampal neurons grown in culture incorporate 3H-uridine in the nucleus, then selectively transport the newly synthesized RNA into dendrites at a rate of about 0.5 mm day-1. The transport is inhibited by metabolic poisons, suggesting that it is an active, energy-dependent process. The RNA may be transported in association with the cytoskeleton.     

FGF-induced vesicular release of Sonic hedgehog and retinoic acid in leftward nodal flow is critical for left-right determination

The precise specification of left-right asymmetry is an essential process for patterning internal organs in vertebrates. In mouse embryonic development, the symmetry-breaking process in left-right determination is initiated by a leftward extraembryonic fluid flow on the surface of the ventral node. However, it is not known whether the signal transduction mechanism of this flow is chemical or mechanical. Here we show that fibroblast growth factor (FGF) signalling triggers secretion of membrane-sheathed objects 0.3-5 microm in diameter termed 'nodal vesicular parcels' (NVPs) that carry Sonic hedgehog and retinoic acid. These NVPs are transported leftward by the fluid flow and eventually fragment close to the left wall of the ventral node. The silencing effects of the FGF-receptor inhibitor SU5402 on NVP secretion and on a downstream rise in Ca2+ were sufficiently reversed by exogenous Sonic hedgehog peptide or retinoic acid, suggesting that FGF-triggered surface accumulation of cargo morphogens may be essential for launching NVPs. Thus, we propose that NVP flow is a new mode of extracellular transport that forms a left-right gradient of morphogens.     

基于双反相二维色谱串联质谱技术的大鼠海马膜蛋白分析

色谱与质谱联用技术在蛋白质组学研究中应用广泛,特别是二维色谱分离技术的发展,为复杂生物样品的分离分析提供了更为精准的技术手段.海马是大脑颞叶内侧的一个重要脑区,主要负责哺乳动物的学习和记忆,实现这些功能的生理过程与海马膜蛋白密切相关,但由于海马膜蛋白具有强疏水性和低丰度的特点,因此在分离和鉴定上难度较高.运用差速离心的方法分离纯化得到成年大鼠的海马膜蛋白,采用双反相二维色谱串联质谱技术进行分离分析.最终鉴定2 502个蛋白,结合生物信息学分析发现,所鉴定到的2 502个蛋白在包含蛋白定位、突触可塑性、蛋白运输以及囊泡转运等在内的与学习记忆功能密切相关的生理过程中均有涉及.这一分析结果为更加全面的揭示海马膜蛋白与脑海马区的特定功能间的具体关系提供了重要的参考.     

N,N′-亚烃基脂肪酸双酰胺的合成及其润滑性能

研究了文题化合物的合成,制备了二十一种该类化合物。红外光谱及元素分析数据证实了这些化合物结构的正确性。由混炼塑化仪测定了它们的润滑性能,探讨了其润滑性能与分子结构之间的关系。结果表明,芥酸的双酰胺较硬脂酸的双酰胺具有更好的润滑作用;特别是N,N′-(1,2-亚乙基)双芥酸酰胺,明显优于国内目前生产的N,N′-(1,2-亚乙基)双硬脂酰胺和芥酸酰胺。     

Two-dimensional spectroscopy of electronic couplings in photosynthesis

Time-resolved optical spectroscopy is widely used to study vibrational and electronic dynamics by monitoring transient changes in excited state populations on a femtosecond timescale. Yet the fundamental cause of electronic and vibrational dynamics--the coupling between the different energy levels involved--is usually inferred only indirectly. Two-dimensional femtosecond infrared spectroscopy based on the heterodyne detection of three-pulse photon echoes has recently allowed the direct mapping of vibrational couplings, yielding transient structural information. Here we extend the approach to the visible range and directly measure electronic couplings in a molecular complex, the Fenna-Matthews-Olson photosynthetic light-harvesting protein. As in all photosynthetic systems, the conversion of light into chemical energy is driven by electronic couplings that ensure the efficient transport of energy from light-capturing antenna pigments to the reaction centre. We monitor this process as a function of time and frequency and show that excitation energy does not simply cascade stepwise down the energy ladder. We find instead distinct energy transport pathways that depend sensitively on the detailed spatial properties of the delocalized excited-state wavefunctions of the whole pigment-protein complex.     

Small-molecule inhibitors of the AAA+ ATPase motor cytoplasmic dynein

The conversion of chemical energy into mechanical force by AAA+ (ATPases associated with diverse cellular activities) ATPases is integral to cellular processes, including DNA replication, protein unfolding, cargo transport and membrane fusion. The AAA+ ATPase motor cytoplasmic dynein regulates ciliary trafficking, mitotic spindle formation and organelle transport, and dissecting its precise functions has been challenging because of its rapid timescale of action and the lack of cell-permeable, chemical modulators. Here we describe the discovery of ciliobrevins, the first specific small-molecule antagonists of cytoplasmic dynein. Ciliobrevins perturb protein trafficking within the primary cilium, leading to their malformation and Hedgehog signalling blockade. Ciliobrevins also prevent spindle pole focusing, kinetochore-microtubule attachment, melanosome aggregation and peroxisome motility in cultured cells. We further demonstrate the ability of ciliobrevins to block dynein-dependent microtubule gliding and ATPase activity in vitro. Ciliobrevins therefore will be useful reagents for studying cellular processes that require this microtubule motor and may guide the development of additional AAA+ ATPase superfamily inhibitors.     

粗糙裂隙溶解过程反应性溶质运移特征

岩体裂隙中溶质运移受对流、扩散和水岩化学反应三种机制控制.分析了不同机制主导作用下，粗糙裂隙溶解过程中反应性溶质运移特征，并分析粗糙度的影响.研究结果显示，粗糙裂隙溶解过程存在4种类型，分别是均匀、非均匀、极度非均匀和均匀-非均匀溶解.其中对流占主导作用条件下，裂隙发生均匀溶解；化学反应占主导作用条件下，流体上游溶解速率高于下游.初始粗糙度对裂隙溶解过程也具有重要影响:初始粗糙度增大会使水岩化学反应速率增大，同时随着水岩反应的进行，裂隙溶解后粗糙度增大.     

有向循环图的强连通度的性质

本文首先证明了连通有向循环图的k原子部分仍为连通有向循环图,并给出了有关连通有向循环图的强连通度的另一些性质。然后利用这些性质得到了基础图是简单图的连通有向循环图的强连通度的下界。     

Molecular cloning and characterization of an insulin-regulatable glucose transporter

A major mechanism by which insulin stimulates glucose transport in muscle and fat is the translocation of glucose transporters from an intracellular membrane pool to the cell surface. The existence of a distinct insulin-regulatable glucose transporter was suggested by the poor cross-reactivity between antibodies specific for either the HepG2 or rat brain glucose transporters and the rat adipocyte glucose transporter. More direct evidence was provided by the production of a monoclonal antibody (mAb 1F8) specific for the rat adipocyte glucose transporter that immunolabels a species of relative molecular mass 43,000 (43K) present only in tissues that exhibit insulin-dependent glucose transport, suggesting that this protein may be encoded by a different gene from the previously described mammalian glucose transporters. This antibody has been used to immunoprecipitate a 43K protein that was photoaffinity-labelled with cytochalasin B in a glucose displaceable way, and to immunolabel a protein in the plasma membrane of rat adipocytes, whose concentration was increased at least fivefold after cellular insulin exposure. Here we describe the cloning and sequencing of cDNAs isolated from both rat adipocyte and heart libraries that encode a protein recognized by mAb 1F8, and which has 65% sequence identity to the human HepG2 glucose transporter. This cDNA hybridizes to an mRNA present only in skeletal muscle, heart and adipose tissue. Our data indicate that this cDNA encodes a membrane protein with the characteristics of the translocatable glucose transporter expressed in insulin-responsive tissues.     

饮料用香精质控体系的研究

介绍了饮料用香精质量控制的一般途径,建立了以感官、理化、成分3个指标相结合的饮料用香精质量控制体系,给出了成分指标检验的实例,并利用夹角余弦法对实例进行估算,得出香精指纹图谱的相似度,以此判断香精质量的稳定性。     

Inhibition of early atherogenesis in transgenic mice by human apolipoprotein AI

Epidemiological surveys have identified a strong inverse relationship between the amount in the plasma of high density lipoproteins (HDL), apolipoprotein AI (ApoA-I), the major protein component of HDL, and the risk for atherosclerosis in humans. It is not known if this relationship arises from a direct antiatherogenic effect of these plasma components or if it is the result of other factors also associated with increases in ApoA-I and HDL levels. Because some strains of mice are susceptible to diet-induced formation of preatherosclerotic fatty streak lesions, and because of available techniques for the genetic manipulation of this organism, the murine system offers a unique setting in which to investigate the process of early atherogenesis. To test the hypothesis that induction of a high plasma concentration of ApoA-I and HDL would inhibit this process, we studied the effects of atherogenic diets on transgenic mice expressing high amounts of human ApoA-I. We report that transgenic mice with high plasma ApoA-I and HDL levels were significantly protected from the development of fatty streak lesions.