Alpha-helical coiled-coil structures of Trypanosoma brucei variable surface glycoproteins

We have used electron microscopy to examine purified intact variable surface glycoproteins (VSGs) from clones derived from two distinct stocks of Trypanosoma brucei. The VSG molecule from MITat 1.2 has a large elongated domain consistent with the shape of the dimeric N-terminal domain determined by X-ray analysis (see preceding paper), and a heretofore unseen short, thin fibrous tail presumed to be the C-terminal domain. Electron microscopy on DiTat 1.3, however, indicates a morphology quite distinct from that of MITat 1.2. Analysis of four VSG amino acid sequences reveals 7-fold periodicities (heptad repeats) which indicate that alpha-helical coiled-coil secondary structure elements occur in all of these VSGs, consistent with the observation of helical bundles in one VSG. These results suggest the possibility that VSG antigenic diversity may be related to a diversity in length and disposition of alpha-helical bundles and coiled-coil domains.     

6 A-resolution X-ray structure of a variable surface glycoprotein from Trypanosoma brucei

The variable surface glycoprotein (VSG) is the predominant component of the surface coat of the African trypanosome. The expression of antigenically distinct VSGs on minor populations during infection allows the parasite to escape the host immune response. Purification of the protein is facilitated by the enzymatic release of a soluble form of VSG (sVSG) which occurs on cell lysis. The soluble form is a dimer with an approximate molecular weight of 120,000-130,000. Partial proteolysis of sVSG reveals a protease-sensitive link between an amino-terminal domain which comprises about two-thirds of the molecule, and a C-terminal domain which contains the membrane attachment site. We have obtained crystals suitable for high-resolution structural analysis from preparations of three sVSG: MITat 1.2, ILTat 1.25 and ILTat 1.22. The crystal structure of the dimer of the MITat 1.2 amino-terminal domain has been solved to 6 A resolution. We report here that the dimer is an unusual 90 A rod-like molecule composed of a helical bundle of at least four 80 A-long alpha-helices.     

Structural organization of the rat thy-1 gene

Thy-1 is a differentiation marker expressed predominantly on thymocytes, T cells and brain tissue. Its presence on murine peripheral T cells but not B cells has long been used to distinguish between these two populations of lymphocytes. Although analogues of Thy-1 have been described in several mammalian species, its tissue distribution in different species varies widely, precluding its use as T-cell-specific marker. The Thy-1 molecule is a cell-surface glycoprotein of relative molecular mass 18,000, one-third of which represents carbohydrate; the protein moieties of the rat and murine Thy-1 molecules have been sequenced and found to consist of 111 and 112 amino acids, respectively. An unusual aspect of Thy-1 is the apparent absence of a hydrophobic segment comparable to that observed in other membrane glycoproteins which would allow integration of Thy-1 within the membrane lipid bilayer. This has prompted speculation that Thy-1 is anchored to the cell surface by some other hydrophobic component such as glycolipid. Here we report the structure of thy-1 complementary DNA and genomic clones and describe the exon-intron organization of the gene. More importantly, our data indicate that Thy-1 is initially synthesized as a molecule of 142 amino acids, 31 amino acids longer at the carboxyl end than the Thy-1 molecule isolated and characterized by Campbell et al. An extremely hydrophobic region of 20 amino acids lies within this 31-amino acid stretch and may represent the transmembrane segment responsible for anchoring Thy-1 to the cell membrane.     

FAST地锚基础工程中的若干问题探讨*

FAST工程主动反射面为一半径300 m、口径500 m的球冠面,是一种基于柔性基底的预应力整体索网结构,索网包括2225个主索节点及下拉索驱动装置和地锚。由于台址岩土单元复杂,地锚点多、分布范围广、定位精度要求高、与场地其他基础设施的相互干涉大,给地锚基础的设计带来了一定的难题。本文针对FAST地锚基础中的特殊问题一一展开研究:以工程地质调查和测绘为基础,通过抓台址区主控因素,集合影响台址岩土性质的自然因素,对岩土单元进行了合理的分区;通过建立台址开挖后的BIM模型,利用极射投影技术实现了地锚中心点的精确定位;利用极射投影结果,对设备之间的相互干涉情况进行了调节和处理。研究成果最终确保了FAST地锚基础工程的顺利竣工。     

全长黏结岩石抗浮锚杆承载性能现场试验

抗浮锚杆具有地层适应能力强、锚固力高、造价低、工期短等优点,具有广阔的工程应用前景.开展了4组13根岩石抗浮锚杆的极限抗拔承载试验,在1根试验锚杆上安装光纤光栅应变传感器进行应力测试,所有试验锚杆均加载至极限破坏状态,从荷载-锚固体顶面位移曲线、锚筋轴力分布、锚筋剪应力分布规律及界面黏结强度等方面进行了分析.结果表明,抗浮锚杆主要出现锚筋-锚固体界面剪切滑移破坏、锚固体-周围岩体界面剪切滑移破坏及锚筋拔断3种破坏形态.试验条件下,黏结长度为2.0 m的抗浮锚杆其极限抗拔承载力为240 kN,黏结长度不小于3.0 m的抗浮锚杆其极限抗拔承载力不低于320 kN,承载力高、变形小,能够满足抗浮要求.锚筋轴力自上而下逐渐衰减,锚筋在距锚固体顶面3.0 m以下范围内不受力,建议中风化花岗岩中抗浮锚杆的黏结长度设计值取3.5～4.0 m.锚筋剪应力沿深度呈先增大后减小的趋势,在距锚固体顶面0.45 m的位置达到峰值,约为2.7 MPa.锚筋-锚固体界面平均黏结强度为1.14～1.36 MPa,锚固体-岩土体界面平均黏结强度为0.28～0.37 MPa.     

螺旋锚垂直抗拔承载力室内试验研究

通过室内螺旋锚上拔模型试验，探讨了单节螺旋锚在砂土和粉土中的埋置深度和锚板直径对其上拔承载力的影响。提出了单节螺旋锚上拔破坏的模式及极限抗拔承载力的计算公式。经过比较，单节螺旋锚的极限抗拔承载力按本文中提出的公式计算，结果与试验实测值比较一致。     

Two variant surface glycoproteins of Trypanosoma brucei of different sequence classes have similar 6 A resolution X-ray structures

Antigenic variation in the African trypanosome is mediated through changes in the composition of the surface coat. By controlling expression of the major surface protein, the variant surface glycoprotein (VSG), from a repertoire of perhaps 1,000 different genes the organisms exhibit a series of antigenically distinct coats and evade the host's immune system. We have determined the 6 A resolution structure of a T. brucei variant surface glycoprotein, ILTat 1.24, using X-ray crystallography. The crystallized protein consists of the N-terminal two-thirds of the intact VSG which has a relative molecular mass (Mr) of 60,000 (60K). The structure, which includes a 90 A long alpha-helical bundle, is strikingly similar to that of the N-terminal fragment of VSG MITat 1.2 (ref. 4). Although most known VSG sequences show little similarity of primary sequence in the N-terminal domain, the similarity between the structure of a Class I (ILTat 1.24) and a Class II (MITat 1.2) VSG antigen suggests that VSGs may share a common tertiary structure.     

混凝土用建筑锚栓拉伸承载力影响因素的试验分析

本文对埋置于混凝土中的建筑锚栓（膨胀摩擦型锚栓）进行了拉伸试验,分析了影响膨胀摩擦型锚栓极限荷载的因素（锚栓的锚杆直径。锚栓的埋深、混凝土的强度）,以及拉伸极限荷载随各因素的变化情况。     

MoFLP1, encoding a novel fungal fasciclin-like protein, is involved in conidiation and pathogenicity in Magnaporthe oryzae

Fasciclin family proteins have been identified as cell adhesion molecules in various organisms. In this study, a novel Magnaporthe oryzae fasciclin-like protein encoding gene, named MoFLP1, was isolated from a subtractive suppressive cDNA library and functionally analyzed. Sequence analysis showed that the MoFLP1 gene contains an open reading frame (ORF) of 1050 nucleotides encoding 349 amino acids with a calculated molecular weight of 35.85 kDa and a pI of 7.76. The deduced MoFLP1 protein contains a 17-amino acid secretion signal sequence and an 18-amino acid sequence with the characteristics of a glycosylphosphotidylinositol (GPI) anchor additional signal at its N- and C-terminuses, respectively. Potential N-glycosylation sites and domains involving cell adhesion were also identified in MoFLP1. Sequence analysis and subcellular localization by the expression of MoFLP1-GFP fusion construct in M. oryzae indicated that the MoFLP1 protein is probably localized on the vacuole membrane. Two MoFLP1 null mutants generated by targeted gene disruption exhibited marked reduction of conidiation, conidial adhesion, appressorium turgor, and pathogenicity. Our results indicate that fasciclin proteins play important roles in fungal development and pathogenicity in M. oryzae.     

Structural and serological evidence for a novel mechanism of antigenic variation in foot-and-mouth disease virus.

Changes resulting in altered antigenic properties of viruses nearly always occur on their surface and have been attributed to the substitution of residues directly involved in binding antibody. To investigate the mechanism of antigenic variation in foot-and-mouth disease virus (FMDV), variants that escape neutralization by a monoclonal antibody have been compared crystallographically and serologically with parental virus. FMDVs form one of the four genera of the Picornaviridae. The unenveloped icosahedral shell comprises 60 copies each of four structural proteins VP1-4. Representatives from each of the genera have similar overall structure, but differences in the external features. For example, human rhinovirus has a pronounced 'canyon' that is proposed to contain the cell attachment site, whereas elements of the attachment site for FMDV, which involves the G-H loop (residues 134-160) and C-terminus (200-213) of VP1, are exposed on the surface. Moreover, this G-H loop, which is a major antigenic site of FMDV, forms a prominent, highly accessible protrusion, a feature not seen in other picornaviruses. It is this loop that is perturbed in the variant viruses that we have studied. The amino acid mutations characterizing the variants are not at positions directly involved in antibody binding, but result in far-reaching perturbations of the surface structure of the virus. Thus, this virus seems to use a novel escape mechanism whereby an induced conformational change in a major antigenic loop destroys the integrity of the epitope.     

外文武围垦工程海堤修复加固结构波浪整体稳定性模型试验

分析了福建省长乐市外文武围垦工程1号海堤护面结构遭受台风破坏的原因,通过波浪整体物理模型试验,研究1号海堤3个特征段海堤修复加固工程外海侧护面结构型式、扭王块体稳定质量与锚系方式,并根据不同水位与波浪组合作用工况下整体模型试验结果,确定了1号海堤修复加固合理方案为镇压层取10 t重扭王块体、规则交错安放且相互锚系的组合型式,坡面为6t重扭王块体、规则安放型式.     

琥珀酰亚胺类固定化酶载体的合成与胺解研究

选用丙烯酸(AA)、N′-羟基琥珀酰亚胺(NHM)为单体,以N′,N′-亚甲基双丙烯酰胺(MBAA)为交联剂,以乙醇水溶液为制孔剂条件下进行反相悬浮聚合,得到珠状共聚物载体,该载体是一种新的酶载体形式,在国内外的相关文献中均未见报道.用红外光谱对其结构进行了表征.对载体与有机胺的胺解反应进行了研究,发现共聚物载体分子链上的丙烯酸-N′-羟基琥珀酰亚胺酯基易于在温和条件下与胺基发生胺解反应,从而为该类载体应用于含游离胺基的生物活性物质分子的固定化提供了依据.     

倾斜螺旋锚片锚杆上拔性能室内试验研究

螺旋锚片锚杆（简称螺旋锚）是一种利用深层土体抗力的锚固结构。笔者通过室内模型试验对螺旋锚的倾斜角度对其上拔承载力的影响及其上拔破坏机理进行了研究和探讨。根据试验结果，利用方差分析得到了一些结论     

锚索应力场分析及其应用研究

针对锚索预应力作用于岩体内形成的应力场状况,利用弹性力学中的Boussinesg解和R.D.Mindlin解,研究处于锚头的岩石是否会由于挤压而产生破坏,锚根附近的岩石是否会由于张拉而发生破坏。通过研究,得出了锚索预应力作用于岩体内各受力部位形成的应力特点;以及为避免群锚效应,锚索应保持的合理间距。将此研究应用于江苏省连云港将军崖岩画的加固工程中,取得了理想的工程效果。     

Targeting of transmembrane and GPI-anchored forms of N-CAM to opposite domains of a polarized epithelial cell.

The calcium-independent neural cell adhesion molecule N-CAM is expressed transiently during development in many tissues, including epithelia. The three naturally occurring principal isoforms of N-CAM differ in the way in which they associate with the membrane and in their cytoplasmic domains. These isoforms are generated by developmentally regulated alternative splicing of a single gene: the large cytoplasmic domain (ld) form (relative molecular mass 180,000 (Mr 180K] is specific for post-mitotic neurons; the 120K small cytoplasmic domain (ssd) and 140K small surface domain (sd) forms also occur on other cell types. One function of the different isoforms could be to specify cellular localization; for example, glycosyl phosphatidyl inositol (GPI)-membrane anchoring acts as a targeting signal for expression on the apical surface of polarized epithelial cells. Neurons and epithelial cells may use similar mechanisms for polarizing their plasma membrane proteins. We have therefore investigated the targeting of GPI-anchored (ssd N-CAM, 120K) and transmembrane forms of N-CAM (sd N-CAM, 140K; ld N-CAM, 180K) by comparing the expression of each after transfection of the appropriate complementary DNAs into polarized epithelial cells. We find that isoforms with alternative modes of membrane association are targeted to different surfaces of polarized epithelial cells: ssd N-CAM is expressed on the apical surface, whereas sd and ld N-CAM are expressed on the basolateral surface. These results suggest that the different isoforms of N-CAM determine their own diverse cellular destinations. They also support the hypothesis that the GPI anchor acts as an apical targeting signal in epithelia.     

新型碳纤维板锚具设计及试验研究

针对传统碳纤维增强复合材料(carbon fiber reinforced plastic, CFRP)板锚具易出现滑脱、劈裂等现象,螺栓提供的预紧力大小难以控制的问题,对传统锚具的结构形式做出改进。通过理论分析新型锚具的受力机理,为新型锚具的结构尺寸设计提供理论依据。利用有限元软件对不同参数CFRP板锚具进行受力模拟,得出锚具设计尺寸进行试验研究。研究结果表明：当夹板横向两侧位移为0.65 mm时,锚具的最大锚固效率为89.37%,CFRP板发生爆炸式破坏;锚具夹板内曲面式设计可通过控制夹板位移有效控制预紧力大小,增强挤压效果。     

组合荷载作用下平板锚承载能力的数值预测

为预测组合荷载作用下平板锚的承载力,假设锚-土之间不脱离,在ABAQUS下建立了法向力、切向力和弯矩共同作用的平板锚运动变形数值模型。与极限理论解对比,证明了上述数值模型的正确,并利用其计算了法向力、切向力和弯矩组合荷载作用下板锚的极限承载力,利用Murff模型拟合了组合荷载作用下板锚的极限承载力包络面。结果表明,Murff模型能较好地拟合组合荷载作用下板锚的极限承载力包络面。     

抗浮锚杆承载性能研究进展

随着中国城市化建设步伐的加快,城市地下空间开发利用已成为热潮,相应的建(构)筑物基坑越来越深、基础埋深不断加大,特别是在地下水丰富且水位较高的地区,结构自身荷载不能有效抵抗地下水浮力时,随之而来的抗浮问题得到了社会的广泛关注。与传统解决建(构)筑物抗浮问题的技术方法相比,抗浮锚杆以单点受力小、地层适应能力强、应力分散、施工速度快、成本低等优点倍受工程界的青睐。对抗浮锚杆的发展历史、锚固性能、应力传递机制等相关研究进行归纳总结,讨论当前抗浮锚杆研究中的不足,并对抗浮锚杆的发展提出建议。     

房产销售人员职业锚类型与工作满意度的关系研究

在Schein职业锚理论的基础上,以房产销售人员为对象,探索职业锚的特征、职业锚与工作满意度的关系以及中介变量。结果显示,房产销售人员的主要职业锚类型是挑战型、创业型和服务型;房产销售人员对同事关系的满意度最高,其次为晋升机会,满意度较低的是报酬;三种主要职业锚类型的房产销售人员的工作满意度水平较高;性别、婚姻状况、年龄阶段、工作年限、学历层次、企业/公司规模等对职业锚类型和工作满意度的各个维度存在不同的影响。研究表明,＂关系＂是职业锚与满意度的中介变量。     

Structure of the membrane-pore-forming fragment of colicin A

Colicins are antibiotic proteins produced by and active against sensitive Escherichia coli and closely related bacteria. They can adsorb to specific receptors located at the external surface of the outer membrane of sensitive cells, and are then translocated to their specific targets within these cells. The largest group of colicins comprises those which can form voltage-dependent channels in membranes, thereby destroying the cell's energy potential. Colicin molecules are organized in structural domains, each domain carrying one function associated with the toxin's lethal activity. The pore-forming activity seems to be located at the carboxyl terminus. A thermolytic fragment comprising amino acids 389-592 from colicin A has pore-forming properties very similar to those of the entire molecule. This fragment is soluble in aqueous medium and spontaneously inserts into lipid bilayers. We have determined the structure of the pore-forming fragment of colicin A by X-ray crystallography and refinement at 2.5 A resolution. The protein consists of ten alpha-helices organized in a three-layer structure. Two of the helices are completely buried within the structure and form a hydrophobic hairpin loop similar to that proposed for signal sequences which function in translocation. We present a model for insertion of the protein into lipid bilayers the features of which may be applicable in other biological systems involving protein insertion or translocation across membranes.