Structural basis of water-specific transport through the AQP1 water channel.

Water channels facilitate the rapid transport of water across cell membranes in response to osmotic gradients. These channels are believed to be involved in many physiological processes that include renal water conservation, neuro-homeostasis, digestion, regulation of body temperature and reproduction. Members of the water channel superfamily have been found in a range of cell types from bacteria to human. In mammals, there are currently 10 families of water channels, referred to as aquaporins (AQP): AQP0-AQP9. Here we report the structure of the aquaporin 1 (AQP1) water channel to 2.2 A resolution. The channel consists of three topological elements, an extracellular and a cytoplasmic vestibule connected by an extended narrow pore or selectivity filter. Within the selectivity filter, four bound waters are localized along three hydrophilic nodes, which punctuate an otherwise extremely hydrophobic pore segment. This unusual combination of a long hydrophobic pore and a minimal number of solute binding sites facilitates rapid water transport. Residues of the constriction region, in particular histidine 182, which is conserved among all known water-specific channels, are critical in establishing water specificity. Our analysis of the AQP1 pore also indicates that the transport of protons through this channel is highly energetically unfavourable.     

Mechanism of ion permeation through calcium channels

Calcium channels carry out vital functions in a wide variety of excitable cells but they also face special challenges. In the medium outside the channel, Ca2+ ions are vastly outnumbered by other ions. Thus, the calcium channel must be extremely selective if it is to allow Ca2+ influx rather than a general cation influx. In fact, calcium channels show a much greater selectivity for Ca2+ than sodium channels do for Na+ despite the high flux that open Ca channels can support. Relatively little is known about the mechanism of ion permeation through Ca channels. Earlier models assumed ion independence or single-ion occupancy. Here we present evidence for a novel hypothesis of ion movement through Ca channels, based on measurements of Ca channel activity at the level of single cells or single channels. Our results indicate that under physiological conditions, the channel is occupied almost continually by one or more Ca2+ ions which, by electrostatic repulsion, guard the channel against permeation by other ions. On the other hand, repulsion between Ca2+ ions allows high throughput rates and tends to prevent saturation with calcium.     

Structural determinants for generating centromeric chromatin

Mammalian centromeres are not defined by a consensus DNA sequence. In all eukaryotes a hallmark of functional centromeres--both normal ones and those formed aberrantly at atypical loci--is the accumulation of centromere protein A (CENP-A), a histone variant that replaces H3 in centromeric nucleosomes. Here we show using deuterium exchange/mass spectrometry coupled with hydrodynamic measures that CENP-A and histone H4 form sub-nucleosomal tetramers that are more compact and conformationally more rigid than the corresponding tetramers of histones H3 and H4. Substitution into histone H3 of the domain of CENP-A responsible for compaction is sufficient to direct it to centromeres. Thus, the centromere-targeting domain of CENP-A confers a unique structural rigidity to the nucleosomes into which it assembles, and is likely to have a role in maintaining centromere identity.     

柔性蒙皮材料氦气渗透的细观机制

该文研究了由聚芳脂纤维(vectran)平纹织物为承力层、PVF膜(tedlar)为阻氦层、聚酯薄膜(mylar)为防护层的浮空器蒙皮材料氦气渗透的细观机制。首先,根据Hagen-Poiseuille定律,推导得到了承力层等效渗透率。然后,通过氦气溶解扩散模型和多层复合结构渗透模型,理论分析了各层对氦气密封性能的贡献比例,讨论了每层不同厚度对整体蒙皮材料氦气渗透率的影响,理论预测了蒙皮材料的整体氦气渗透率。最后,将实验结果与理论所得的数值进行了对比,验证了多层渗透模型的正确性。     

Ion permeation through the Na+,K+-ATPase

P-type ATPase pumps generate concentration gradients of cations across membranes in nearly all cells. They provide a polar transmembrane pathway, to which access is strictly controlled by coupled gates that are constrained to open alternately, thereby enabling thermodynamically uphill ion transport (for example, see ref. 1). Here we examine the ion pathway through the Na+,K+-ATPase, a representative P-type pump, after uncoupling its extra- and intracellular gates with the marine toxin palytoxin. We use small hydrophilic thiol-specific reagents as extracellular probes and we monitor their reactions, and the consequences, with cysteine residues introduced along the anticipated cation pathway through the pump. The distinct effects of differently charged reagents indicate that a wide outer vestibule penetrates deep into the Na+,K+-ATPase, where the pathway narrows and leads to a charge-selectivity filter. Acidic residues in this region, which are conserved to coordinate pumped ions, allow the approach of cations but exclude anions. Reversing the charge at just one of those positions converts the pathway from cation selective to anion selective. Close structural homology among the catalytic subunits of Ca2+-, Na+,K+- and H+,K+-ATPases argues that their extracytosolic cation exchange pathways all share these physical characteristics.     

河流水资源结构分析研究

针对河流中水体的功能和作用,从生态保护、兴利运用和防洪减灾的角度出发,提出了河流水资源结构分析的思想和方法.河流水资源可划分为生态水量、安全水量、风险水量和灾害水量四部分,各部分水量由特定的流量来界定.以北方河流为背景,在对4种水量的概念及其功能与作用深入分析的基础上,提出了各类水量的计算方法;研究了松花江哈尔滨水文断面的水资源结构划分和各类水量分配规律.结果表明,所提出的方法可以对河流水资源进行定量分析,进而实现分类管理、高效利用.     

Structural determinants for GoLoco-induced inhibition of nucleotide release by Galpha subunits

Heterotrimeric G-proteins bind to cell-surface receptors and are integral in transmission of signals from outside the cell. Upon activation of the Galpha subunit by binding of GTP, the Galpha and Gbetagamma subunits dissociate and interact with effector proteins for signal transduction. Regulatory proteins with the 19-amino-acid GoLoco motif can bind to Galpha subunits and maintain G-protein subunit dissociation in the absence of Galpha activation. Here we describe the structural determinants of GoLoco activity as revealed by the crystal structure of Galpha(i1) GDP bound to the GoLoco region of the 'regulator of G-protein signalling' protein RGS14. Key contacts are described between the GoLoco motif and Galpha protein, including the extension of GoLoco's highly conserved Asp/Glu-Gln-Arg triad into the nucleotide-binding pocket of Galpha to make direct contact with the GDP alpha- and beta-phosphates. The structural organization of the GoLoco Galpha(i1) complex, when combined with supporting data from domain-swapping experiments, suggests that the Galpha all-helical domain and GoLoco-region carboxy-terminal residues control the specificity of GoLoco Galpha interactions.     

Impairment of angiogenesis and cell migration by targeted aquaporin-1 gene disruption

Aquaporin-1 (AQP1) is a water channel protein expressed widely in vascular endothelia, where it increases cell membrane water permeability. The role of AQP1 in endothelial cell function is unknown. Here we show remarkably impaired tumour growth in AQP1-null mice after subcutaneous or intracranial tumour cell implantation, with reduced tumour vascularity and extensive necrosis. A new mechanism for the impaired angiogenesis was established from cell culture studies. Although adhesion and proliferation were similar in primary cultures of aortic endothelia from wild-type and from AQP1-null mice, cell migration was greatly impaired in AQP1-deficient cells, with abnormal vessel formation in vitro. Stable transfection of non-endothelial cells with AQP1 or with a structurally different water-selective transporter (AQP4) accelerated cell migration and wound healing in vitro. Motile AQP1-expressing cells had prominent membrane ruffles at the leading edge with polarization of AQP1 protein to lamellipodia, where rapid water fluxes occur. Our findings support a fundamental role of water channels in cell migration, which is central to diverse biological phenomena including angiogenesis, wound healing, tumour spread and organ regeneration.     

冷冻法净化苦咸水的实验

利用调温冰柜在-4℃,-8℃,-10℃和-12℃的条件下对祖厉河水样进行实验研究,实验结果表明:冷冻法对苦咸水中超标离子(阴阳离子)的脱盐效果较显著;碎冰过滤可以进一步提高冰晶质量,使各超标离子平均总脱盐率都在95%以上;脱盐效果受冷冻温度和溶液质量浓度的影响,随着冷冻温度的降低或苦咸水质量浓度的增加呈现下降趋势.通过研究,可以充分利用甘肃省冬季的气候条件,丰富的咸水资源,采用冷冻法来淡化本地区的苦咸水,从而解决祖厉河流域上的饮水问题.     

Expression of aquaporin-1 in SMMC-7221 liver carcinoma cells promotes cell migration

Aquaporins (AQPs) are membrane water channels that play pivotal roles in physiological and pathophysi- ological processes in diverse mammalian organs[1―3]. Recent studies indicated a novel role of AQPs in cell migration. Mice lacking AQP1, the endothelia…     

Structural determinants for regulation of phosphodiesterase by a G protein at 2.0 A

A multitude of heptahelical receptors use heterotrimeric G proteins to transduce signals to specific effector target molecules. The G protein transducin, Gt, couples photon-activated rhodopsin with the effector cyclic GMP phosophodiesterase (PDE) in the vertebrate phototransduction cascade. The interactions of the Gt alpha-subunit (alpha(t)) with the inhibitory PDE gamma-subunit (PDEgamma) are central to effector activation, and also enhance visual recovery in cooperation with the GTPase-activating protein regulator of G-protein signalling (RGS)-9 (refs 1-3). Here we describe the crystal structure at 2.0 A of rod transducin alpha x GDP x AlF4- in complex with the effector molecule PDEgamma and the GTPase-activating protein RGS9. In addition, we present the independently solved crystal structures of the RGS9 RGS domain both alone and in complex with alpha(t/i1) x GDP x AlF4-. These structures reveal insights into effector activation, synergistic GTPase acceleration, RGS9 specificity and RGS activity. Effector binding to a nucleotide-dependent site on alpha(t) sequesters PDEgamma residues implicated in PDE inhibition, and potentiates recruitment of RGS9 for hydrolytic transition state stabilization and concomitant signal termination.     

Structural relaxation in supercooled water by time-resolved spectroscopy

Water has many kinetic and thermodynamic properties that exhibit an anomalous dependence on temperature, in particular in the supercooled phase. These anomalies have long been interpreted in terms of underlying structural causes, and their experimental characterization points to the existence of a singularity at a temperature of about 225 K. Further insights into the nature and origin of this singularity might be gained by completely characterizing the structural relaxation in supercooled water. But until now, such a characterization has only been realized in simulations that agree with the predictions of simple mode-coupling theory; unambiguous experimental support for this surprising conclusion is, however, not yet available. Here we report time-resolved optical Kerr effect measurements that unambiguously demonstrate that the structural relaxation of liquid and weakly supercooled water follows the behaviour predicted by simple mode-coupling theory. Our findings thus support the interpretation of the singularity as a purely dynamical transition. That is, the anomalous behaviour of weakly supercooled water can be explained using a fully dynamic model and without needing to invoke a thermodynamic origin. In this regard, water behaves like many other, normal molecular liquids that are fragile glass-formers.     

低欠热水通过渐扩喷嘴的泄放研究

旨在通过实验确定５ＭＷ核供热堆注硼系统所用渐扩喷管的水力特性。研究了在压力为０．７～１．５ＭＰａ的范围内，欠热度约为２０℃的水通过喉部直径为２～８ｍｍ、扩张角为６０°～１２０°的渐扩喷嘴的流动情况，结果表明：在一定条件下可在这种扩张角的渐扩喷嘴中形成临界流动现象。当出现临界流动时，通过渐扩喷嘴的临界流量大于孔板临界流量公式的计算值；当在喷嘴中没有形成临界流动时，通过这种渐扩喷嘴的流量可用孔板流量公式计算；流体通过扩张角为９０°的大扩张角渐扩喷嘴时的流动损失小于通过孔板时的损失；随扩张角的增大，相同条件下通过喷嘴的流量减小。     

Biodiversity improves water quality through niche partitioning

Excessive nutrient loading of water bodies is a leading cause of water pollution worldwide, and controlling nutrient levels in watersheds is a primary objective of most environmental policy. Over the past two decades, much research has shown that ecosystems with more species are more efficient at removing nutrients from soil and water than are ecosystems with fewer species. This has led some to suggest that conservation of biodiversity might be a useful tool for managing nutrient uptake and storage, but this suggestion has been controversial, in part because the specific biological mechanisms by which species diversity influences nutrient uptake have not been identified. Here I use a model system of stream biofilms to show that niche partitioning among species of algae can increase the uptake and storage of nitrate, a nutrient pollutant of global concern. I manipulated the number of species of algae growing in the biofilms of 150 stream mesocosms that had been set up to mimic the variety of flow habitats and disturbance regimes that are typical of natural streams. Nitrogen uptake rates, as measured by using (15)N-labelled nitrate, increased linearly with species richness and were driven by niche differences among species. As different forms of algae came to dominate each unique habitat in a stream, the more diverse communities achieved a higher biomass and greater (15)N uptake. When these niche opportunities were experimentally removed by making all of the habitats in a stream uniform, diversity did not influence nitrogen uptake, and biofilms collapsed to a single dominant species. These results provide direct evidence that communities with more species take greater advantage of the niche opportunities in an environment, and this allows diverse systems to capture a greater proportion of biologically available resources such as nitrogen. One implication is that biodiversity may help to buffer natural ecosystems against the ecological impacts of nutrient pollution.     

Structural basis for semaphorin signalling through the plexin receptor

Semaphorins and their receptor plexins constitute a pleiotropic cell-signalling system that is used in a wide variety of biological processes, and both protein families have been implicated in numerous human diseases. The binding of soluble or membrane-anchored semaphorins to the membrane-distal region of the plexin ectodomain activates plexin's intrinsic GTPase-activating protein (GAP) at the cytoplasmic region, ultimately modulating cellular adhesion behaviour. However, the structural mechanism underlying the receptor activation remains largely unknown. Here we report the crystal structures of the semaphorin 6A (Sema6A) receptor-binding fragment and the plexin A2 (PlxnA2) ligand-binding fragment in both their pre-signalling (that is, before binding) and signalling (after complex formation) states. Before binding, the Sema6A ectodomain was in the expected 'face-to-face' homodimer arrangement, similar to that adopted by Sema3A and Sema4D, whereas PlxnA2 was in an unexpected 'head-on' homodimer arrangement. In contrast, the structure of the Sema6A-PlxnA2 signalling complex revealed a 2:2 heterotetramer in which the two PlxnA2 monomers dissociated from one another and docked onto the top face of the Sema6A homodimer using the same interface as the head-on homodimer, indicating that plexins undergo 'partner exchange'. Cell-based activity measurements using mutant ligands/receptors confirmed that the Sema6A face-to-face dimer arrangement is physiologically relevant and is maintained throughout signalling events. Thus, homodimer-to-heterodimer transitions of cell-surface plexin that result in a specific orientation of its molecular axis relative to the membrane may constitute the structural mechanism by which the ligand-binding 'signal' is transmitted to the cytoplasmic region, inducing GAP domain rearrangements and activation.     

建立清洁开发机制促进经济结构调整

阐述了CDM（清洁开发机制）的含义与作用,分析了CDM对山西解决环境污染问题的重要性及实现的可能性,指出在山西建立CDM,是有效改善山西环境和搞好经济结构调整的最佳途径。     

低渗透岩石的单相水非Darcy流

为研究低渗透岩石的非 Darcy渗流现象 ,采用FDES- 6 4 1三轴驱替评价系统进行了稳定渗流实验 ,测定了盐水和蒸馏水在低渗透砂岩和砾岩岩样中的渗流特性。岩样抽真空后水平放置于岩样夹持器中 ,通过多级驱替泵控制密闭岩样中水的渗流速度 ,所产生的压力差由压力传感器测定。结论为 :1)在空气渗透率非常低并且孔隙度非常小的砾岩中 ,明显存在着非 Darcy渗流现象 ,但由渗流曲线后退趋势所得出的启动压力梯度非常小 ,以致于在生产应用中可以忽略 ;2 )低渗透介质中的微粒运移是低速非 Darcy渗流现象存在的主要原因 ;3)岩石的非均匀性降低了岩石的渗透率。     

锅炉恒差压供水控制系统

利用变频技术控制水泵的转速,同时利用电子技术对锅炉的汽压信号和供水信号进行处理,控制锅炉供水,达到节能和安全的目的,该系统在印染厂十吨锅炉上试用后,效果非常明显。