ABEEMσπ/MM模型对蛋白质G_A88/G_B88水溶液动力学性质的研究

应用ABEEMσπ/MM（σπ水平的原子与键电负性均衡方法融合进分子力学）浮动电荷模型以及显性ABEEM-7P水模型,对GA88和GB88两个蛋白质分子进行了分子动力学模拟.分析了2个蛋白质的动力学性质,包括蛋白质的回旋半径、疏水表面积和亲水表面积、各类原子位置的均方根偏差以及氢键分布.通过对比水溶液和真空下2个蛋白质的回旋半径,表明该模型很好地体现了蛋白质的＂电致紧缩＂现象;对疏水表面积和亲水表面积的计算表明,GB88中残基与溶剂的相互作用更强一些;非氢原子位置的均方根偏差及氢键分布情况与实验结构相比较表明,ABEEMσπ/MM浮动电荷模型模拟的GA88和GB88的结构与实验结构有很好的一致性,进而说明该模型的合理性和参数的可转移性.     

Topographical rearrangement of acetylcholine receptors alters channel kinetics

Plasma membranes are dynamic structures of proteins and lipids. Protein-protein or protein-lipid interactions within the membrane are believed to have important roles in many membrane functions, including ion transport, enzyme activity and signal reception. The acetylcholine (ACh) receptor-channel complex in skeletal muscle membrane is one of the best known integral membrane proteins. Its ion transport function is accessible to direct measurement at the single-channel level by the use of the 'giga-seal' patch recording technique. Here we used an in situ electrophoresis technique to rearrange the topography of pre-existing ACh receptor-channels in the muscle membrane, and measured the single-channel kinetics of ACh-activated channels in two different molecular environments within the membrane: those in the diffusely distributed region and those in the ACh receptor clusters induced by the applied field. We found that the channel kinetics are significantly prolonged in the ACh receptor cluster compared with the non-clustered region of the same cell. This result strongly supports the notion that the function of a membrane ionic channel depends on the local molecular environment.     

Ser-His-Glu triad forms the catalytic site of the lipase from Geotrichum candidum

The Ser-His-Asp triad is a well known structural feature of the serine proteases. It has also been directly observed in the catalytic sites of two lipases, whose high-resolution three-dimensional structures have been determined 1,2. Lipases show a wide variety of sizes, substrate and positional specificities, and catalytic rates 3. They achieve maximal catalytic rates at oil-water interfaces. The fungus Geotrichum candidum produces several different forms of lipases, two of which have been purified to homogeneity 4,5. Two lipase genes have been identified, cloned and sequenced 6,7. Both code for proteins of 544 amino acids with a total relative molecular mass of about 60,000 (Mr 60K). The two forms are 86% identical. Their isoelectric points differ slightly, being between 4.3 and 4.6. About 7% of the total Mr is carbohydrate. Until now, only a low resolution structure of GCL has been reported 8, but no high resolution structure has followed. We now report the three-dimensional structure of a lipase from G. candidum (GCL) at 2.2 A resolution. Unlike the other lipases and serine proteases, the catalytic triad of GCL is Ser-His-Glu, with glutamic acid replacing the usual aspartate. Although the sequence similarity with the other two lipases is limited to the region near the active-site serine, there is some similarity in their three-dimensional structures. The GCL is also an alpha/beta protein with a central mixed beta sheet whose topology is similar to that of the N-terminal domain of human pancreatic lipase. As in the other lipases 1,2, the catalytic site is buried under surface loops. Sequence comparisons with proteins from the cholinesterase family suggest that they also contain the Ser-His-Glu triad.     

酶法改性对蛋白质结构性质的影响及与酶特异性的关系

采用分子生物物理学原理探讨了蛋白质的平均分子量、水化面积、极性、憎水性、柔性及电荷分布与酶作用的关系.研究表明:改性蛋白质的平均分子量与水解度成反比例关系;水化面积随水解度的增加成对数关系增大;极性与平均分子量成反比例关系;柔性与平均分子量成负指数函数关系;净电荷数在pH、温度和蛋白质浓度一定的条件下,随水解度的增加成正比例关系增大.改性蛋白质的憎水性由两部分构成,分别为肽键断裂引起憎水性的变化和构象变化引起的变化.其中肽键断裂引起憎水性变化与水解度成正比关系.总之,改性蛋白质的结构性质变化与酶作用之间存在明显的定量关系,而且结构性质的变化还与酶特异性有关.     

Homologous plant and bacterial proteins chaperone oligomeric protein assembly

An abundant chloroplast protein is implicated in the assembly of the oligomeric enzyme ribulose bisphosphate carboxylase-oxygenase, which catalyses photosynthetic CO2-fixation in higher plants. The product of the Escherichia coli groEL gene is essential for cell viability and is required for the assembly of bacteriophage capsids. Sequencing of the groEL gene and the complementary cDNA encoding the chloroplast protein has revealed that these proteins are evolutionary homologues which we term 'chaperonins'. Chaperonins comprise a class of molecular chaperones that are found in chloroplasts, mitochondria and prokaryotes. Assisted post-translational assembly of oligomeric protein structures is emerging as a general cellular phenomenon.     

低聚炔基芴衍生物电子光谱的理论研究

运用密度泛函(DFT)B3LYP方法、从头算单激发组态相互作用(CIS)方法分别优化了炔基取代芴的单体和三聚体的基态及最低激发单重态几何结构.系统分析了分子结构、前线分子轨道特征以探索电子跃迁机理.应用含时密度泛函理论(TD-DFT)计算了分子的电子光谱,得到三聚芴炔的最大吸收及发射光谱分别为417和447 nm,与实验结果接近.同时探讨了单体和低聚体系不同的发光特征,指出炔基芴的低聚体系是良好的蓝光材料.     

大地电磁测深法探测山区深埋隧道隐伏构造——以安石隧道探测为例

隧道掌子面前方与上方存在的隐伏构造在隧道施工过程中具有极强的致灾性.针对云南凤庆安石隧道具有长、大、深埋,且其地表起伏大的特点,采用大地电磁测深法对隧道上方隐伏构造进行探测.首先对观测数据进行静态效应校正等预处理;随后进行带地形的基于光滑约束最小二乘反演,得到了隧道轴线及平行轴线的视电阻率剖面图;最后利用反演结果并结合研究区的区域地质构造、岩性特征以及水文地质条件等,揭示出与隧道斜交的两条隐伏构造的空间展布,该成果与后期钻探结果吻合,表明大地电磁测深法可以有效地探测出山区深埋隧道前方与上方隐伏构造的位置与姿态,为隧道安全施工与地质灾害防治提供了重要的参考依据.     

Transient association of newly synthesized unfolded proteins with the heat-shock GroEL protein

It has been suggested that newly synthesized proteins are maintained in their unfolded state by cellular ATP-driven factors which may prevent or reverse the formation of misfolded structures or promote the correct assembly of oligomeric proteins or post-translational secretion. Using a photocross-linking approach, we have identified the 20S heat-shock GroEL protein as the major cytosolic component which forms a complex with the unfolded newly synthesized pre-beta-lactamase or chloramphenicol acetyltransferase in Escherichia coli. Dissociation of these complexes is ATP-dependent. The unfolded state of pre-beta-lactamase, maintained by the transient interaction with GroEL, may be essential for the secretion of this protein.     

Subtractive proteomic mapping of the endothelial surface in lung and solid tumours for tissue-specific therapy

The molecular complexity of tissues and the inaccessibility of most cells within a tissue limit the discovery of key targets for tissue-specific delivery of therapeutic and imaging agents in vivo. Here, we describe a hypothesis-driven, systems biology approach to identifying a small subset of proteins induced at the tissue-blood interface that are inherently accessible to antibodies injected intravenously. We use subcellular fractionation, subtractive proteomics and bioinformatics to identify endothelial cell surface proteins exhibiting restricted tissue distribution and apparent tissue modulation. Expression profiling and gamma-scintigraphic imaging with antibodies establishes two of these proteins, aminopeptidase-P and annexin A1, as selective in vivo targets for antibodies in lungs and solid tumours, respectively. Radio-immunotherapy to annexin A1 destroys tumours and increases animal survival. This analytical strategy can map tissue- and disease-specific expression of endothelial cell surface proteins to uncover novel accessible targets useful for imaging and therapy.     

Evolutionary conservation of biogenesis of beta-barrel membrane proteins

The outer membranes of mitochondria and chloroplasts are distinguished by the presence of beta-barrel membrane proteins. The outer membrane of Gram-negative bacteria also harbours beta-barrel proteins. In mitochondria these proteins fulfil a variety of functions such as transport of small molecules (porin/VDAC), translocation of proteins (Tom40) and regulation of mitochondrial morphology (Mdm10). These proteins are encoded by the nucleus, synthesized in the cytosol, targeted to mitochondria as chaperone-bound species, recognized by the translocase of the outer membrane, and then inserted into the outer membrane where they assemble into functional oligomers. Whereas some knowledge has been accumulated on the pathways of insertion of proteins that span cellular membranes with alpha-helical segments, very little is known about how beta-barrel proteins are integrated into lipid bilayers and assembled into oligomeric structures. Here we describe a protein complex that is essential for the topogenesis of mitochondrial outer membrane beta-barrel proteins (TOB). We present evidence that important elements of the topogenesis of beta-barrel membrane proteins have been conserved during the evolution of mitochondria from endosymbiotic bacterial ancestors.     

聚苯硫醚的合成与结构研究I.聚苯硫醚的合成与结构表征

用不同原料和方法制备了不同类型的聚苯硫醚树脂.通过元素分析、红外光谱、激光拉曼光谱及X射线衍射分析等手段,对各种产品的结构进行了表征.对不同类型产品的结构差异进行了初步研究.结果表明,用不同的原料和方法都可制备聚苯硫醚树脂,各种聚苯硫醚树脂的结构基本接近,但具有细微差别.     

四川盆地超深层泥页岩纳米孔隙特征及其地质意义

利用SEM-PCAS孔隙定量表征技术与低压N_2等温吸附实验研究X井深度为6 875~8 042m超深层泥页岩的纳米孔隙特征,并选取四川盆地及周缘地区从地表到5km左右的样品组作为对比,探索深埋藏作用对泥页岩孔隙系统的影响。研究发现,X井志留系龙马溪组、奥陶系五峰组及下寒武统筇竹寺组超深层泥页岩32个样品的孔隙特征相似,孔隙类型以有机质孔、粒间孔为主,孔隙形态以狭长-裂缝型为主。N_2等温吸附线类型为IV-H3型,QSDFT孔径分布显示其纳米孔隙主要分布于4~16nm段,BET比表面积为8.63~16.13m2/g。与对照组样品相比,X井超深层泥页岩的孔径分布更加分散,微孔体积和微孔比表面积更小,介孔/微孔的体积比值及介孔/微孔的比表面积比值比非超深层泥页岩均具有数量级的优势。X井超深层泥页岩的孔隙特征主要受埋藏深度控制,深埋藏作用会使泥页岩孔径缩小并改变孔隙的形态。     

聚苯硫醚的合成与结构研究：Ⅰ.聚苯硫醚的合成与结构表征

用不同原料和方法制备了不同类型的聚苯硫醚树脂,通过元素分析。红外光谱,激光拉曼光谱及Ｘ射线衍射分析等手段,对各种产品的结构进行了表征。对不同类型产品的结构差异进行了初步研究。结果表明,用不同的原料和方法都可制备聚苯硫醚树脂,各种聚苯硫醚树脂的结构基本接近,但具有细微差别。     

寒区油气集输管道合适埋深数值计算

油气集输管道的敷设深度直接影响管道的建设成本,对管道的安全平稳运行也起重要作用.考虑寒区土壤的初始温度周期性变化、地表不同日平均温度以及是否冻结等因素,建立数学模型,并采用有限单元法对多种埋设深度的集输管道土壤温度场进行数值计算.实例计算表明:0.8 m是寒区较合适的管道埋设深度.当大气温度升高时,可适当减小集输管道的埋深,以节省建设成本.在集输管道运行管理计算分析时,可以不考虑冻结及非冻结状态物性对土壤自然温度场的影响,将土壤看作常物性.该模型可为寒区油气集输管道的敷设提供一定的理论依据.     

东营凹陷王古1潜山构造特征与含油气性分析

王古1潜山是首次在东营凹陷钻探成功的深部潜山.通过对王古1潜山的构造特征、形成演化过程研究,认为王古1潜山属于内幕式的褶皱块断山,是经历了从古生代到新生代多期次挤压、拉张应力场相互叠加所发展起来的潜山,其内幕逆冲推覆构造为断层传播褶皱.在此基础上,探讨了王古1潜山的烃源岩、储集层以及油气运移通道等条件,最后,提出在该地区下一步勘探的有利区域.     

Inhibition of antigen-induced T-cell clone proliferation by antigen-specific antibodies

Attempts to inhibit the recognition of soluble antigens by T lymphocytes using antibodies specific for the antigen in question have been uniformally unsuccessful, in contrast to the observed specific inhibition of antibody generation by B cells. One exception is the unique situation whereby anti-hapten antisera inhibit the T-cell proliferative responses observed when hapten-specific T lymphocytes or clones are cultured with hapten-derivatized cells or proteins. The inability to inhibit T-cell functions by antigen-specific antibodies has been interpreted in several ways: (1) T cells possess a different repertoire from B cells; (2) the antibodies tested recognize epitopes present on the native antigen, whereas T cells recognize non-native (processed) structures; (3) the antigenic determinant(s) recognized by T cells on the surface of antigen presenting cells are either not accessible to antibodies, or are present in low amounts. The development of antigen-specific T-cell clones and monoclonal antibodies both specific for the same antigenic determinants now allows this question to be investigated definitively. Here, we report for the first time the specific inhibition of antigen-induced T-cell clone proliferation by a monoclonal antibody directed against the relevant soluble protein antigen.     

大庆长垣南部黑帝庙油层浅层气特征

大庆长垣南部黑帝庙油层发育于嫩江组第三段、第四段地层中,属松辽盆地上部含油组合.研究认为,本地区的天然气分布主要受构造和沉积控制,断层对天然气的运移起到至关重要的作用,古构造的高点地区(现在处于斜坡带)是现今天然气的有利聚集区;本区沉积相主要为三角洲前缘亚相,砂体厚度相对较小,模糊识别天然气预测显示该油层天然气呈分散状分布;但总体看,由于本区天然气埋藏很浅,容易开发,且天然气预测分布面积之总和具一定规模,因此有较好的发展前景.     

羊毛溶解再利用的研究现状

角蛋白质是生物组织结构中一种重要的结构蛋白质，是天然的高分子物。其分子结构又杂不易溶解。现有研究，主要是利用废弃羊毛来制备角蛋白质溶液，希望得到的溶液角蛋白质分子不受或少受破坏，并保持长链结构。虽然已有很多溶解方法，但真正生态、无损溶解较少，溶解对角蛋白分子量影响的解释就更少。通过对已有研究和利用的回顾，以提出存在的问题和研究方向。     

聚氯乙烯中烯丙基氯含量的研究

用电位滴定法测定了聚氯乙烯(PVC) 中的烯丙基氯含量, 研究了烯丙基氯含量与PVC 相对分子质量的关系; 用氯仿萃取了PVC 中不同相对分子质量的级分, 测定了二个级分的烯丙基氯含量。结果发现聚合物中较低相对分子质量部分的炳丙基氯含量比较高相对分子质量部分的高。表明在聚合反应中控制相对分子质量分布是很重要的, 为了提高PVC 的热稳定性, 在氯乙烯聚合反应中要减少低相对分子质量部分的PVC 的生成。     

Quantitative dynamics and binding studies of the 20S proteasome by NMR

The machinery used by the cell to perform essential biological processes is made up of large molecular assemblies. One such complex, the proteasome, is the central molecular machine for removal of damaged and misfolded proteins from the cell. Here we show that for the 670-kilodalton 20S proteasome core particle it is possible to overcome the molecular weight limitations that have traditionally hampered quantitative nuclear magnetic resonance (NMR) spectroscopy studies of such large systems. This is achieved by using an isotope labelling scheme where isoleucine, leucine and valine methyls are protonated in an otherwise highly deuterated background in concert with experiments that preserve the lifetimes of the resulting NMR signals. The methodology has been applied to the 20S core particle to reveal functionally important motions and interactions by recording spectra on complexes with molecular weights of up to a megadalton. Our results establish that NMR spectroscopy can provide detailed insight into supra-molecular structures over an order of magnitude larger than those routinely studied using methodology that is generally applicable.