Structural Basis for Dimerization and Catalysis of a Novel Esterase from the GTSAG Motif Subfamily of the Bacterial Hormone-sensitive Lipase Family

Hormone-sensitive lipases (HSLs) are widely distributed in microorganisms, plants, and animals. Microbial HSLs are classified into two subfamilies, an unnamed new subfamily and the GDSAG motif subfamily. Due to the lack of structural information, the detailed catalytic mechanism of the new subfamily is not yet clarified. Based on sequence analysis, we propose to name the new subfamily as the GTSAG motif subfamily. We identified a novel HSL esterase E25, a member of the GTSAG motif subfamily, by functional metagenomic screening, and resolved its structure at 2.05 Å. E25 is mesophilic (optimum temperature at 50 °C), salt-tolerant, slightly alkaline (optimum pH at 8.5) for its activity, and capable of hydrolyzing short chain monoesters (C2–C10). E25 tends to form dimers both in the crystal and in solution. An E25 monomer contains an N-terminal CAP domain, and a classical α/β hydrolase-fold domain. Residues Ser¹⁸⁶, Asp²⁸², and His³¹² comprise the catalytic triad. Structural and mutational analyses indicated that E25 adopts a dimerization pattern distinct from other HSLs. E25 dimer is mainly stabilized by an N-terminal loop intersection from the CAP domains and hydrogen bonds and salt bridges involving seven highly conserved hydrophilic residues from the catalytic domains. Further analysis indicated that E25 also has some catalytic profiles different from other HSLs. Dimerization is essential for E25 to exert its catalytic activity by keeping the accurate orientation of the catalytic Asp²⁸² within the catalytic triad. Our results reveal the structural basis for dimerization and catalysis of an esterase from the GTSAG motif subfamily of the HSL family.     

Immune protection induced on day 10 following administration of the 2009 A/H1N1 pandemic influenza vaccine

Background

The 2009 swine-origin influenza virus (S-OIV) H1N1 pandemic has caused more than 18,000 deaths worldwide. Vaccines against the 2009 A/H1N1 influenza virus are useful for preventing infection and controlling the pandemic. The kinetics of the immune response following vaccination with the 2009 A/H1N1 influenza vaccine need further investigation.

Methodology/Principal Findings

58 volunteers were vaccinated with a 2009 A/H1N1 pandemic influenza monovalent split-virus vaccine (15 µg, single-dose). The sera were collected before Day 0 (pre-vaccination) and on Days 3, 5, 10, 14, 21, 30, 45 and 60 post vaccination. Specific antibody responses induced by the vaccination were analyzed using hemagglutination inhibition (HI) assay and enzyme-linked immunosorbent assay (ELISA). After administration of the 2009 A/H1N1 influenza vaccine, specific and protective antibody response with a major subtype of IgG was sufficiently developed as early as Day 10 (seroprotection rate: 93%). This specific antibody response could maintain for at least 60 days without significant reduction. Antibody response induced by the 2009 A/H1N1 influenza vaccine could not render protection against seasonal H1N1 influenza (seroconversion rate: 3% on Day 21). However, volunteers with higher pre-existing seasonal influenza antibody levels (pre-vaccination HI titer ≥1∶40, Group 1) more easily developed a strong antibody protection effect against the 2009 A/H1N1 influenza vaccine as compared with those showing lower pre-existing seasonal influenza antibody levels (pre-vaccination HI titer <1∶40, Group 2). The titer of the specific antibody against the 2009 A/H1N1 influenza was much higher in Group 1 (geometric mean titer: 146 on Day 21) than that in Group 2 (geometric mean titer: 70 on Day 21).

Conclusions/Significance

Recipients could gain sufficient protection as early as 10 days after vaccine administration. The protection could last at least 60 days. Individuals with a stronger pre-existing seasonal influenza antibody response may have a relatively higher potential for developing a stronger humoral immune response after vaccination with the 2009 A/H1N1 pandemic influenza vaccine.     

Effects of Cerium on Key Enzymes of Carbon Assimilation of Spinach Under Magnesium Deficiency

The mechanism of the fact that cerium improves the photosynthesis of plants under magnesium deficiency is poorly understood. The main aim of the study was to determine the role of cerium in the amelioration of magnesium deficiency effects in CO₂ assimilation of spinach. Spinach plants were cultivated in Hoagland’s solution. They were subjected to magnesium deficiency and to cerium chloride administered in the magnesium-present Hoagland’s media and magnesium-deficient Hoagland’s media. The results showed that the chlorophyll synthesis and oxygen evolution was destroyed, and the activities of Rubisco carboxylasae and Rubisco activase and the expression of Rubisco large subunit (rbcL), Rubisco small subunit (rbcS), and Rubisco activase subunit (rca) were significantly inhibited, then plant growth was inhibited by magnesium deficiency. However, cerium promotes the chlorophyll synthesis, the activities of two key enzymes in CO₂ assimilation, and the expression of rbcL, rbcS, and rca, thus leading to the enhancement of spinach growth under magnesium-deficient conditions.     

蒙古扁桃的开花生物学研究

对分布于宁夏的蒙古扁桃居群进行开花动态、繁育系统、花粉与柱头活性、生殖器官解剖结构与花粉管的生长观察以及人工授粉实验,分析其传粉生物学特性,以探讨影响蒙古扁桃结实率低下的原因.结果显示:蒙古扁桃为专性异交植物;花具有花柱多型性现象,同一个体中存在长花柱花、中花柱花、短花柱花3种花柱型花,在不同个体中三型花所占比例不同且差异显著;短花柱花不可育,中花柱花与长花柱花为可育花.研究表明,大量不育花的存在、早春稀少的传粉昆虫以及荒漠地区恶劣的气候环境和破碎化的生境是导致蒙古扁桃结实率低下的主要因素.     

不同剂量^137Cs-γ辐射对毛竹幼苗叶片叶绿素荧光参数的影响

利用不同剂量的137Cs-γ射线对毛竹（Phyllostachys heterocycla ‘Pubescens’）种子进行辐射, 测定实生苗叶片中的光合色素含量和叶绿素荧光参数等指标, 探讨辐射对毛竹幼苗生长的影响, 为筛选有利的突变单株奠定良好基础。结果表明：30或60 Gy的137Cs-γ射线辐射后, 毛竹幼苗的光合色素含量以及最大荧光强度（Fm）、可变荧光强度（Fv）、PSII最大光化学效率（Fv/Fm）、PSII的潜在活性（Fv/Fo）、PSII实际光化学效率（Yield）和表观光合电子传递速率（ETR）等荧光参数值均高于90 Gy辐射处理, 说明较低剂量辐射后PSII反应中心的能量捕获效率高, 且具有较强的光合能力; 而90 Gy的137Cs-γ射线辐射对毛竹的影响则与之相反。不同处理剂量之间叶片光能耗散程度以及表观光合电子传递速率-光合有效辐射（ETR-PAR）响应曲线的分析结果也进一步证实了以上结论。     

线粒体内膜解偶联蛋白UCP1在大肠杆菌中的活性表达

线粒体的呼吸耗氧偶联着ATP的合成,而位于线粒体内膜上的跨膜蛋白解偶联蛋白(uncoupling protein,UCP)能够破坏这种偶联关系.在大肠杆菌中表达了有生物活性的鼠源解偶联蛋白1(rUCP1).重组rUCP1的表达导致大肠杆菌宿主细胞生长变慢;在电子显微镜下观察免疫标记的结果显示,重组rUCP1主要表达在细菌膜上;同时将rUCP1重构到脂质体中也能够测到质子转运活性.这些结果说明,真核生物UCP1能够在原核生物中表达出有生物活性的形式,且能纯化得到足量的rUCP1蛋白用于进一步的结构生物学研究.     

LISS-DF近端螺钉不同固定方式的有限元分析

目的研究LISS-DF治疗股骨远端骨折近端螺钉不同固定方式对应力分布规律的影响,寻找符合生物力学原理的固定方式,为临床应用提供理论依据。方法在ANSYS9.0软件中建立LISS-DF钢板固定股骨远端骨折（AO/OTA33-A3型）的实体模型和有限元模型。在近端螺钉不同单、双皮质固定方式下,通过模拟生理应力做轴向加载实验并进行有限元计算和分析。结果1、3孔单2、4孔双皮质固定时,近端螺钉最大应力值在不同固定方式中最小为24.21975Mpa,位移值为0.131424um与其它固定方式相当且均较小,说明该固定方式可以取得较好的力学效果。结论近端螺钉靠近骨折端处双皮质固定,其余螺钉间隔单双皮质固定时,可以减少应力集中现象,得到更好的把持力,使固定更牢固,从而降低固定失败的风险性。     

Brain-derived Neurotrophic Factor (BDNF)-induced Mitochondrial Motility Arrest and Presynaptic Docking Contribute to BDNF-enhanced Synaptic Transmission

Appropriate mitochondrial transport and distribution are essential for neurons because of the high energy and Ca²⁺ buffering requirements at synapses. Brain-derived neurotrophic factor (BDNF) plays an essential role in regulating synaptic transmission and plasticity. However, whether and how BDNF can regulate mitochondrial transport and distribution are still unclear. Here, we find that in cultured hippocampal neurons, application of BDNF for 15 min decreased the percentage of moving mitochondria in axons, a process dependent on the activation of the TrkB receptor and its downstream PI3K and phospholipase-Cγ signaling pathways. Moreover, the BDNF-induced mitochondrial stopping requires the activation of transient receptor potential canonical 3 and 6 (TRPC3 and TRPC6) channels and elevated intracellular Ca²⁺ levels. The Ca²⁺ sensor Miro1 plays an important role in this process. Finally, the BDNF-induced mitochondrial stopping leads to the accumulation of more mitochondria at presynaptic sites. Mutant Miro1 lacking the ability to bind Ca²⁺ prevents BDNF-induced mitochondrial presynaptic accumulation and synaptic transmission, suggesting that Miro1-mediated mitochondrial motility is involved in BDNF-induced mitochondrial presynaptic docking and neurotransmission. Together, these data suggest that mitochondrial transport and distribution play essential roles in BDNF-mediated synaptic transmission.     

孔雀石绿对中华大蟾蜍蝌蚪的毒性研究

孔雀石绿(ＭａｌａｃｈｉｔｅＧｒｅｅｎ)是一种三苯甲烷染料,由于它有很强的毒性,多年来在水产养殖的疾病防治中经常使用:在鱼类养殖中使用015～02ｐｐｍ的浓度全池遍洒或用10ｐｐｍ的浓度浸洗15～30分钟来防治鱼类的水霉病等(郑曙明,1997)。同时用0002～0005ｐｐｍ孔雀石绿全池泼洒,或用02～05ｐｐｍ药浴2小时,或6～7ｐｐｍ药浴5～10分钟,也常用于防治蛙类蝌蚪的水霉病等(吴高升,1997)。孔雀绿直接进入水环境作为一种杀菌剂在水产养殖中使用多年,但对两栖类蝌蚪的影响末见报道,本文采用生物学上常用的实验动物中华大蟾蜍(Ｂｕｆｏｇａｒｇ…