Modulation of folding pathways of exported proteins by the leader sequence

Leader peptides that function to direct export of proteins through membranes have some common features but exhibit a remarkable sequence diversity. Thus there is some question whether leader peptides exert their function through conventional stereospecific protein-protein interaction. Here it is shown that the leader peptides retarded the folding of precursor maltose-binding protein and ribose-binding protein from Escherichia coli. This kinetic effect may be crucial in allowing precursors to enter the export pathway.     

No specific recognition of leader peptide by SecB, a chaperone involved in protein export

Most proteins destined for export from Escherichia coli are made as precursors containing amino-terminal leader sequences that are essential for export and that are removed during the process. The initial step in export of a subset of proteins, which includes maltose-binding protein, is binding of the precursor by the molecular chaperone SecB. This work shows directly that SecB binds with high affinity to unfolded maltose-binding protein but does not specifically recognize and bind the leader. Rather, the leader modulates folding to expose elements in the remainder of the polypeptide that are recognized by SecB.     

Role of the ABC transporter Mdl1 in peptide export from mitochondria

ATP-binding cassette (ABC) adenosine triphosphatases actively transport a wide variety of compounds across biological membranes. Here, the ABC protein Mdl1 was identified as an intracellular peptide transporter localized in the inner membrane of yeast mitochondria. Mdl1 was required for mitochondrial export of peptides with molecular masses of approximately 2100 to 600 daltons generated by proteolysis of inner-membrane proteins by the m-AAA protease in the mitochondrial matrix. Proteolysis by the i-AAA protease in the intermembrane space led to the release of similar-sized peptides independent of Mdl1. Thus, two pathways of peptide efflux from mitochondria exist that may allow communication between mitochondria and their cellular environment.     

Multiple mechanisms of protein insertion into and across membranes

Protein localization in cells is initiated by the binding of characteristic leader (signal) peptides to specific receptors on the membranes of mitochondria or endoplasmic reticulum or, in bacteria, to the plasma membrane. There are differences in the timing of protein synthesis and translocation into or across the bilayer and in the requirement for a transmembrane electrochemical potential. Comparisons of protein localization in these different membranes suggest underlying common mechanisms.     

Unusual pattern of accumulation of mRNA encoding EGF-related protein in sea urchin embryos

A sea urchin (Strongylocentrotus purpuratus) messenger RNA encoding a protein (SpEGF2) related to epidermal growth factor (EGF) was identified. The full-length complementary DNA sequence predicts a protein with an unusually simple structure, including four tandem EGF-like repeats and a hydrophobic leader, but lacking a potential transmembrane domain. Sequence similarities suggest that the peptides are homologous to two peptides from a different sea urchin species, which cause a classic developmental defect, exogastrulation, when added to the seawater outside of embryos. The SpEGF2 messenger RNA begins to accumulate at blastula stage, and in pluteus larvae it is distributed in discrete regions of ectoderm that are not congruent with known histological borders. One region corresponds to that expressing the homeodomain-containing protein, SpHbox1. The structure of the SpEGF2 protein and the pattern of accumulation of its messenger RNA suggest that it may have important functions as a secreted factor during development of sea urchin embryos.     

The function of a leader peptide in translocating charged amino acyl residues across a membrane

Insertion of bacteriophage coat proteins into the membrane of infected bacterial cells can be studied as a model system of protein translocation across membranes. The coat protein of the filamentous bacteriophage Pf3--which infects Pseudomonas aeruginosa--is 44 amino acids in length and has the same basic structure as the coat protein of bacteriophage M13, which infects Escherichia coli. However, unlike the Pf3 coat protein, the M13 coat protein is synthesized as a precursor (procoat) with a typical leader (signal) sequence, which is cleaved after membrane insertion. Nevertheless, when the gene encoding the Pf3 coat protein is expressed in E. coli, the protein is translocated across the membrane. Hybrid M13 and Pf3 coat proteins were constructed in an attempt to understand how the Pf3 coat protein is translocated without a leader sequence. These studies demonstrated that the extracellular regions of the proteins determined their cellular location. When three charged residues in this region were neutralized, the leader-free M13 coat protein was also inserted into the membrane. Differences in the water shell surrounding these residues may account for efficient membrane insertion of the protein without a leader sequence.     

Proton motive force involved in protein transport across the outer membrane of Aeromonas salmonicida

Many Gram-negative bacteria export proteins to the exterior. Some of these proteins are first secreted into the periplasm and then cross the outer membrane in a separate step. The source of energy required for the translocation is unknown. Export of the extracellular protein proaerolysin from the periplasm through the outer membrane of Aeromonas salmonicida is inhibited by a proton ionophore and by low extracellular pH. One possible explanation of these results is that a proton gradient across the outer membrane is required for export.     

The 35-nucleotide spliced leader sequence is common to all trypanosome messenger RNA's

In Trypanosomatidae the messenger RNA's (mRNA's) that code for the variant surface glycoproteins (VSG's), tubulins, calmodulin, and at least a subset of other proteins contain a common 35-nucleotide leader sequence at their 5' ends. Hybrid-arrested in vitro translation has been used to show that all mRNA's in both African and South American trypanosomes contain this 35-nucleotide sequence. Oligonucleotides complementary to this sequence blocked translation of all trypanosome mRNA's in a rabbit reticulocyte lysate system, but did not inhibit translation of mRNA's from other organisms lacking this sequence. An oligonucleotide complementary to the VSG mRNA downstream from the spliced leader sequence arrested only VSG synthesis. Thus, the 35-nucleotide leader sequence is a general feature of all trypanosome mRNA's. The high specificity of oligonucleotides complementary to the spliced leader for their target sequence suggests that analogues permeable to the cell membrane may be useful in the treatment of trypanosomal infections.     

Bacteriophage M13 procoat protein inserts into the plasma membrane as a loop structure

The major coat protein of bacteriophage M13 is synthesized as a precursor, the procoat, with a typical leader (signal) sequence of 23 residues at its NH2-terminus. A fusion protein that contains the NH2-terminal 141 residues of cytoplasmic ribulokinase and all but the first ten residues of M13 procoat was made. The fusion protein inserts into the plasma membrane of Escherichia coli and is processed by leader peptidase to give rise to a leader peptide of 155 residues and the mature coat protein of 50 residues. The NH2-terminus of the leader peptide remains in the cytoplasm and is protected from protease added to the medium outside of the cell. This indicates that M13 procoat inserts into the membrane as a loop structure and that the NH2-terminus of a leader peptide remains within the cytoplasm during membrane insertion.     

云南省农产品出口增长影响因素分析——基于CMS模型的实证研究

运用CMS模型分析了云南省2001～2010年农产品出口增加的成因,结果表明,世界农产品进口规模的扩大、产品出口竞争力的提高以及产品出口结构适应世界进口需求结构的变化是推动云南省农产品出口增加的主要因素,而且近几年农产品出口竞争力的提高成为云南省农产品出口增长的关键因素;从分类产品上看,较强的出口竞争力促进植物性产品和饮料、烟草等产品的出口,而较弱的出口竞争力以及不合理的出口结构阻碍了动物性产品和动植物油脂产品的出口。     

Vertical Flux of Biogenic Carbon in the Ocean: Is There Food Web Control?

Models of biogenic carbon (BC) flux assume that short herbivorous food chains lead to high export, whereas complex microbial or omnivorous food webs lead to recycling and low export, and that export of BC from the euphotic zone equals new production (NP). In the Gulf of St. Lawrence, particulate organic carbon fluxes were similar during the spring phytoplankton bloom, when herbivory dominated, and during nonbloom conditions, when microbial and omnivorous food webs dominated. In contrast, NP was 1.2 to 161 times greater during the bloom than after it. Thus, neither food web structure nor NP can predict the magnitude or patterns of BC export, particularly on time scales over which the ocean is in nonequilibrium conditions.     

安徽省农产品出口现状与提升策略研究

分析了2006—2015年安徽省农产品出口现状:总体态势良好,特产出口占主导,出口组织化程度提升,投资成为出口新渠道;但相对规模仍然偏小,产品附加值低,潜力未能充分挖掘。为做大做强安徽省农产品出口贸易,必须依托丰富的自然资源,打造特色产业,延伸产业链条,提高加工深度,强化品牌地位,全面提升出口竞争力。     

水分胁迫对水稻籽粒灌浆及淀粉合成有关酶活性的影响

 【目的】探索不同水稻抗旱品种在水分胁迫条件下籽粒灌浆特性及有关淀粉酶活性的变化规律。【方法】选用抗旱性不同的3个杂交水稻品种,分别在正常供水和水分胁迫条件下,研究水稻茎鞘物质运转、籽粒灌浆和淀粉合成有关酶活性。【结果】水分胁迫下,抽穗期和成熟期茎鞘干物重以及抽穗后茎鞘物质输出量、输出率、转化率均下降;籽粒生长潜势、最大灌浆速率和平均灌浆速率降低,籽粒生长活跃期缩短,各品种达最大灌浆速率的时间提前;籽粒灌浆过程中可溶性淀粉合成酶（SSS）、Q酶和ADPG焦磷酸化酶3种酶活性都不同程度的被抑制。各性状抗旱性强的品种均比抗旱性弱的品种减少的幅度小。抽穗期茎鞘干物重、抽穗后茎鞘干物质输出量、输出率和转化率均与籽粒产量呈显著或极显著正相关。【结论】在水分胁迫下抗旱性强的品种具有较高的茎鞘物质输出率和转化率、籽粒灌浆速率及淀粉合成有关酶活性,是产量降低较少的生理基础。     

Vertical nitrate fluxes in the oligotrophic ocean

The vertical flux of nitrate across the thermocline in the upper ocean imposes a rigorous constraint on the rate of export of organic carbon from the surface layer of the sea. This export is the primary means by which the oceans can serve as a sink for atmospheric carbon dioxide. For the oligotrophic open ocean regions, which make up more than 75% of the world's ocean, the rate of export is currently uncertain by an order of magnitude. For most of the year, the vertical flux of nitrate is that due to vertical turbulent transport of deep water rich in nitrate into the relatively impoverished surface layer. Direct measurements of rates of turbulent kinetic energy dissipation, coupled with highly resolved vertical profiles of nitrate and density in the oligotrophic eastern Atlantic showed that the rate of transport, averaged over 2 weeks, was 0.14 (0.002 to 0.89, 95% confidence interval) millimole of nitrate per square meter per day and was statistically no different from the integrated rate of nitrate uptake as measured by incorporation of (15)N-labeled nitrate. The stoichiometrically equivalent loss of carbon from the upper ocean, which is the relevant quantity for the carbon dioxide and climate question, is then fixed at 0.90 (0.01 to 5.70) millimole of carbon per square meter per day. These rates are much lower than recent estimates based on in situ changes in oxygen over annual scales; they are consistent with a biologically unproductive oligotrophic ocean.     

Small phytoplankton and carbon export from the surface ocean

Autotrophic picoplankton dominate primary production over large oceanic regions but are believed to contribute relatively little to carbon export from surface layers. Using analyses of data from the equatorial Pacific Ocean and Arabian Sea, we show that the relative direct and indirect contribution of picoplankton to export is proportional to their total net primary production, despite their small size. We suggest that all primary producers, not just the large cells, can contribute to export from the surface layer of the ocean at rates proportional to their production rates.     

Increase in the export of alkalinity from North America's largest river

Chemical weathering and the subsequent export of carbonate alkalinity (HCO3- + CO3-2) from soils to rivers account for significant amounts of terrestrially sequestered atmospheric CO2. We show here that during the past half-century, the export of this alkalinity has increased dramatically from North America's largest river, the Mississippi. This increased export is in part the result of increased flow resulting from higher rainfall in the Mississippi basin. Subcatchment data from the Mississippi suggest that the increase in the export of alkalinity is also linked to amount and type of land cover. These observations have important implications for the potential management of carbon sequestration in the United States.     

山东省蜂蜜出口竞争力比较分析

根据2004～2010年统计数据,介绍了山东省蜂蜜出口状况,并对我国几个蜂蜜出口大省的蜂蜜出口量和出口单价进行了比较,指出山东省蜂蜜出口量的竞争力逐步增强,但不具备价格竞争优势.最后利用国际市场占有率和显示性比较优势指数对山东省蜂蜜出口竞争力进行了测算和分析,结果认为山东省蜂蜜出口具有显示性比较优势,很有希望进一步扩大国际市场.     

FDI直接出口效应和间接出口效应——基于区域视角的实证研究

将外商直接投资对出口的促进作用细分为直接出口效应和间接出口效应,以1994-2007年统计数据为基础,通过构建出口导向指数,分析外商直接投资企业出口发展趋势;运用动态面板GMM模型估计FDI对内资企业间接出口效应。结果显示：外资企业出口导向趋势逐渐降低,内销份额逐渐扩大;FDI对内资企业出口具有显著正的间接效应,但弹性较低,即FDI对内资企业出口的促进作用不很明显。     

小麦蓝矮植原体免疫膜蛋白（Imp）基因的克隆和分子特性分析

 【目的】探讨小麦蓝矮植原体免疫膜蛋白在介体-病原-寄主互作的分子机理。【方法】通过植原体免疫膜蛋白基因序列两侧的保守区设计引物对Imp 1051/Imp 2265,用PCR方法扩增小麦蓝矮植原体免疫膜蛋白基因;对扩增片段的最大开放阅读框和基因的同源矩阵、系统发育树分析;对克隆基因所编码蛋白进行跨膜区、亲疏水区和前导信号序列分析。【结果】从小麦蓝矮病病株和接种长春花中均扩增到约1.0 kb的特异片段,其中小麦蓝矮植原体免疫膜蛋白基因长495 bp,推导的编码蛋白含有164个氨基酸。与10种植原体的免疫膜蛋白基因进行序列同源性分析,小麦蓝矮与三叶草绿变植原体同源性最高,核苷酸和编码的氨基酸序列的同源率分别为98.4％和95.1％。蛋白质结构分析结果表明：小麦蓝矮免疫膜蛋白N端有一个跨膜的前导信号序列,C端为跨膜锚定区,中间为膜外亲水区。【结论】小麦蓝矮植原体与三叶草绿变、翠菊黄化、洋葱黄化和泡桐丛枝植原体的免疫膜蛋白为同型蛋白。     

出口贸易风险测度及其影响因素分析——基于跨国数据的经验研究

将部门的出口份额与构建的部门产出方差协方差矩阵相结合，通过定义一国出口结构风险指数，并利用出口结构风险指数对样本国家在样本期内的出口风险进行测算。同时，把出口风险指数分解为赫芬达尔项、平均风险项、协方差效应项以及赫芬达尔项与平均风险项的交互项。最后，选取影响一国出口风险的主要宏观经济变量，利用跨国面板数据对出口风险进行影响因素分析，结果表明：在诸多宏观经济因素中，出口贸易条件和金融开放程度对一国出口风险的影响最为显著，而一国的人均收入水平则对该国的出口风险影响不明显。