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1.
We sought to develop nuclear DNA (nDNA) probes which could be used to complement mtDNA and DNA fingerprinting markers in distinguishing striped bass, Morone saxatilis (Walbaum), from discrete spawning systems. Restriction endonuclease-generated single copy, 10–20-kb striped bass nuclear nDNA fragments were cloned into the bacteriophage vector Lambda Dash II and tested in Southern blot analyses for their abilities to reveal population-specific polymorphisms. Three of the I7 nDNA sequences tested exhibited polymorphisms which potentially could be used to delineate striped bass populations. One probe, DSB 22, revealed significant genotypic frequency differences between Gulf of Mexico and Atlantic striped bass and among striped bass representative of some Atlantic systems. These preliminary results suggest that single copy nDNA sequences may provide sufficient polymorphisms to aid in stock identification of species which proved genetically monomorphic using other approaches. 相似文献
2.
Mutual information and entropy transfer analysis employed on two inactive states of human beta-2 adrenergic receptor (β2-AR) unraveled distinct communication pathways. Previously, a so-called “highly” inactive state of the receptor was observed during 1.5 microsecond long molecular dynamics simulation where the largest intracellular loop (ICL3) was swiftly packed onto the G-protein binding cavity, becoming entirely inaccessible. Mutual information quantifying the degree of correspondence between backbone-Cα fluctuations was mostly shared between intra- and extra-cellular loop regions in the original inactive state, but shifted to entirely different regions in this latest inactive state. Interestingly, the largest amount of mutual information was always shared among the mobile regions. Irrespective of the conformational state, polar residues always contributed more to mutual information than hydrophobic residues, and also the number of polar-polar residue pairs shared the highest degree of mutual information compared to those incorporating hydrophobic residues. Entropy transfer, quantifying the correspondence between backbone-Cα fluctuations at different timesteps, revealed a distinctive pathway directed from the extracellular site toward intracellular portions in this recently exposed inactive state for which the direction of information flow was the reverse of that observed in the original inactive state where the mobile ICL3 and its intracellular surroundings drove the future fluctuations of extracellular regions. 相似文献
3.
Nevena Cvetesic Mirna Bilus Ita Gruic-Sovulj 《The Journal of biological chemistry》2015,290(22):13981-13991
Aminoacyl-tRNA synthetases catalyze ATP-dependent covalent coupling of cognate amino acids and tRNAs for ribosomal protein synthesis. Escherichia coli isoleucyl-tRNA synthetase (IleRS) exploits both the tRNA-dependent pre- and post-transfer editing pathways to minimize errors in translation. However, the molecular mechanisms by which tRNAIle organizes the synthetic site to enhance pre-transfer editing, an idiosyncratic feature of IleRS, remains elusive. Here we show that tRNAIle affects both the synthetic and editing reactions localized within the IleRS synthetic site. In a complex with cognate tRNA, IleRS exhibits a 10-fold faster aminoacyl-AMP hydrolysis and a 10-fold drop in amino acid affinity relative to the free enzyme. Remarkably, the specificity against non-cognate valine was not improved by the presence of tRNA in either of these processes. Instead, amino acid specificity is determined by the protein component per se, whereas the tRNA promotes catalytic performance of the synthetic site, bringing about less error-prone and kinetically optimized isoleucyl-tRNAIle synthesis under cellular conditions. Finally, the extent to which tRNAIle modulates activation and pre-transfer editing is independent of the intactness of its 3′-end. This finding decouples aminoacylation and pre-transfer editing within the IleRS synthetic site and further demonstrates that the A76 hydroxyl groups participate in post-transfer editing only. The data are consistent with a model whereby the 3′-end of the tRNA remains free to sample different positions within the IleRS·tRNA complex, whereas the fine-tuning of the synthetic site is attained via conformational rearrangement of the enzyme through the interactions with the remaining parts of the tRNA body. 相似文献
4.
Niyati Jain Christopher E. Morgan Brittany D. Rife Marco Salemi Blanton S. Tolbert 《The Journal of biological chemistry》2016,291(5):2331-2344
Splicing patterns in human immunodeficiency virus type 1 (HIV-1) are maintained through cis regulatory elements that recruit antagonistic host RNA-binding proteins. The activity of the 3′ acceptor site A7 is tightly regulated through a complex network of an intronic splicing silencer (ISS), a bipartite exonic splicing silencer (ESS3a/b), and an exonic splicing enhancer (ESE3). Because HIV-1 splicing depends on protein-RNA interactions, it is important to know the tertiary structures surrounding the splice sites. Herein, we present the NMR solution structure of the phylogenetically conserved ISS stem loop. ISS adopts a stable structure consisting of conserved UG wobble pairs, a folded 2X2 (GU/UA) internal loop, a UU bulge, and a flexible AGUGA apical loop. Calorimetric and biochemical titrations indicate that the UP1 domain of heterogeneous nuclear ribonucleoprotein A1 binds the ISS apical loop site-specifically and with nanomolar affinity. Collectively, this work provides additional insights into how HIV-1 uses a conserved RNA structure to commandeer a host RNA-binding protein. 相似文献
5.
Boundary layer properties of highly dissected leaves: an investigation using an electrochemical fluid tunnel 总被引:2,自引:0,他引:2
Abstract. A method for modelling heat and mass transfer by diffusion-controlled electrode reactions in a fluid tunnel is described. In this procedure, a nickelplated leaf functions as a test electrode, and the convective transfer of ions to the leaf cathode in an electrolyte-filled flow tunnel is measured as a function of flow rate. The method permits the simulation of water vapour and heat transfer, and in particular, the determination of boundary layer conductances, by analogy with observed ion transfer. The approach is applicable to many problems in modelling heat and mass transfer between leaves and their surroundings, and is especially useful in examining the properties of leaves in which surface characteristics or overall shape are complex. Using this method, the properties of the highly dissected leaves of Achillea lanulosa with regard to forced convection were investigated. The leaves showed high transfer conductances, indicating that the effective unit of heat transfer was probably the individual leaf subelements. Conductances tended to be greater and effective characteristic dimensions smaller for the larger, more open leaves of a lower altitude population in contrast with leaves from high altitude plants. While the results provide insight into the properties of these complex leaf shapes, difficulties in interpreting the findings are discussed, and a number of exploratory approaches are suggested for data analysis and interpretation. 相似文献
6.
中国梧桐属(Firmiana)在世界梧桐属中占比较大,且除梧桐外其余种均为中国特有且分布范围狭窄的植物种,灭绝风险大,研究气候变化对中国梧桐属树种的影响对于维护生物多样性具有重要的意义。结合多时期第六次国际气候耦合模式比较计划(CMIP6)气候变量数据和中国八种梧桐属树种的分布数据,基于R语言kuenm程序包优化的最大熵(Maxent)模型模拟分析中国八种梧桐属树种在多尺度下的潜在适生区,得出梧桐属最适宜的模拟尺度、潜在适生区的面积变化和迁移方向、梧桐属多样性保护关键区域及保护空缺。结果表明:(1)梧桐属最适宜的模拟尺度为亚洲;(2) Maxent模型的接收者操作特征曲线下面积(AUC)值均大于0.9,表明模型对梧桐属潜在适生区预测结果具有较高准确度;(3)气候变化影响下除云南梧桐(Firmiana major)外其它树种的潜在适生区都将在未来有所扩大;(4)中国八种梧桐属树种潜在适生区迁移方向主要为东西向,南北向大跨度迁移较少,纬度变化不大;(5)丹霞梧桐(Firmiana danxiaensis)的稳定潜在适生区最小;(6)中国梧桐属多样性保护关键区域主要分布于广西壮族自治区及云南、广东、海南等省区;(7)中国梧桐属多样性保护空缺区域主要分布于广西壮族自治区中部及海南省北部;(8)梧桐属多样性保护关键区域正在为人造地表所侵蚀。研究分析气候变化对中国八种梧桐属树种的影响及其潜在适生区变化、中国梧桐属多样性保护状态,可为中国梧桐属建立多样性保护廊道提供相关建议,为制定多样性保护规划及相应措施提供参考。 相似文献
7.
8.
《Bioorganic & medicinal chemistry》2020,28(9):115437
NDM-1 can hydrolyze nearly all available β-lactam antibiotics, including carbapenems. NDM-1 producing bacterial strains are worldwide threats. It is still very challenging to find a potent NDM-1 inhibitor for clinical use. In our study, we used a metal-binding pharmacophore (MBP) enriched virtual fragment library to screen NDM-1 hits. SPR screening helped to verify the MBP virtual hits and identified a new NDM-1 binder and weak inhibitor A1. A solution NMR study of 15N-labeled NDM-1 showed that A1 disturbed all three residues coordinating the second zinc ion (Zn2) in the active pocket of NDM-1. The perturbation only happened in the presence of zinc ion, indicating that A1 bound to Zn2. Based on the scaffold of A1, we designed and synthesized a series of NDM-1 inhibitors. Several compounds showed synergistic antibacterial activity with meropenem against NDM-1 producing K. pneumoniae. 相似文献
9.
10.
《Chirality》2017,29(11):716-725
The absolute configuration (AC) of the naturally occurring ocimenes (−)‐(3S ,5Z )‐2,6‐dimethyl‐2,3‐epoxyocta‐5,7‐diene ( 1 ) and (−)‐(3S ,5Z )‐2,6‐dimethylocta‐5,7‐dien‐2,3‐diol ( 2 ), isolated from the essential oils of domesticated specimens of Artemisia absinthium , followed by vibrational circular dichroism (VCD) studies of 1 , as well as from the acetonide 3 and the monoacetate 4 , both derived from 2 , since secondary alcohols are not the best functional groups to be present during VCD studies in solution due to intermolecular associations. The AC follows from comparison of experimental and calculated VCD spectra that were obtained by Density Functional Theory computation at the B3LYP/DGDZVP level of theory. Careful nuclear magnetic resonance (NMR) measurements were compared with literature values, providing for the first time systematic 1H and 13C chemical shift data. Regarding homonuclear 1H coupling constants, after performing a few irradiation experiments that showed the presence of several small long‐range interactions, the complete set of coupling constants for 3 , which is representative of the four studied molecules, was determined by iterations using the PERCH software. This procedure even allowed assigning the pro ‐R and pro ‐S methyl group signals of the two gem ‐dimethyl groups present in 3 . 相似文献