Reconstitution of the mitochondrial PrxIII antioxidant defence pathway: general properties and factors affecting PrxIII activity and oligomeric state

The mitochondrial 2-Cys peroxiredoxin PrxIII serves as a thioredoxin-dependent peroxidase operating in tandem with its cognate partners, an organelle-specific thioredoxin (Trx2) and NADP-linked thioredoxin reductase (TRR2). This PrxIII pathway is emerging as a primary regulator of intracellular H(2)O(2) levels with dual roles in antioxidant defence and H(2)O(2)-mediated signalling. Here we describe the reconstitution of the mammalian PrxIII pathway in vitro from its purified recombinant components and investigate some of its overall properties. Employing the site-directed PrxIII mutants C47S, C66S and C168S, the putative N and C-terminal catalytic cysteine residues are shown to be essential for function whereas the C66S mutant retains full activity. The pathway attains maximal capacity at low H(2)O(2) concentrations (<10 microM) and is progressively inhibited in the range 0.1 mM to 1.0 mM peroxide. Damage to PrxIII caused by over-oxidation is confirmed by the appearance of abnormal oxidised species of PrxIII on SDS-PAGE at elevated H(2)O(2) levels. The presence of an N-terminal His-tag on PrxIII markedly enhances dodecamer stability, particularly apparent in its oxidised state. Its removal promotes oxidised PrxIII dissociation into dimers and leads to a 3.0-3.5-fold stimulation in peroxidase activity. The unusual concatenated crystal structure of PrxIII consisting of two-interlocked dodecameric rings is also evident in dilute solution employing transmission electron microscopy; however, it represents only 3-5% of the population with most molecules present as single toroids. Moreover, concatenated PrxIII C168S reverts to single toroids on crystal dissolution indicating that these higher-order structures are produced dynamically during the crystallisation process.     

Expression changes of hypothalamic Ahi1 in mice brain: implication in sensing insulin signaling

Growing evidence suggests that the brain, in particular the hypothalamus, directly senses hormones and nutrients to initiate feeding behavior and metabolic responses in the control of energy homeostasis. However, the molecular mechanisms underlying this important process have remained largely unknown. Our study provides the evidence for the role of Abelson helper integration site 1 (Ahi1) protein as a sensor of insulin signaling in the hypothalamus. We found that fasting increased the expression of hypothalamic Ahi1 which was accompanied by lower levels of circulating insulin compared with satiated mice, while re-feeding decreased the expression of hypothalamic Ahi1 which was accompanied by higher levels of circulating insulin. We also found the up-regulated expression of hypothalamic Ahi1 in high-fat induced obese mice, db/db mice, and streptozotocin induced diabetic mice. In addition, we demonstrated that insulin could decrease the expression of Ahi1 in neuroblastoma cell line N18TG2. Taken together, our results indicate that hypothalamic Ahi1 functions as a sensor of insulin signaling.     

Dolichol phosphate mannose synthase: a Glycosyltransferase with Unity in molecular diversities

N-glycans provide structural and functional stability to asparagine-linked (N-linked) glycoproteins, and add flexibility. Glycan biosynthesis is elaborative, multi-compartmental and involves many glycosyltransferases. Failure to assemble N-glycans leads to phenotypic changes developing infection, cancer, congenital disorders of glycosylation (CDGs) among others. Biosynthesis of N-glycans begins at the endoplasmic reticulum (ER) with the assembly of dolichol-linked tetra-decasaccharide (Glc₃Man₉GlcNAc₂-PP-Dol) where dolichol phosphate mannose synthase (DPMS) plays a central role. DPMS is also essential for GPI anchor biosynthesis as well as for O- and C-mannosylation of proteins in yeast and in mammalian cells. DPMS has been purified from several sources and its gene has been cloned from 39 species (e.g., from protozoan parasite to human). It is an inverting GT-A folded enzyme and classified as GT2 by CAZy (carbohydrate active enZyme; http://www.cazy.org). The sequence alignment detects the presence of a metal binding DAD signature in DPMS from all 39 species but finds cAMP-dependent protein phosphorylation motif (PKA motif) in only 38 species. DPMS also has hydrophobic region(s). Hydropathy analysis of amino acid sequences from bovine, human, S. crevisiae and A. thaliana DPMS show PKA motif is present between the hydrophobic domains. The location of PKA motif as well as the hydrophobic domain(s) in the DPMS sequence vary from species to species. For example, the domain(s) could be located at the center or more towards the C-terminus. Irrespective of their catalytic similarity, the DNA sequence, the amino acid identity, and the lack of a stretch of hydrophobic amino acid residues at the C-terminus, DPMS is still classified as Type I and Type II enzyme. Because of an apparent bio-sensing ability, extracellular signaling and microenvironment regulate DPMS catalytic activity. In this review, we highlight some important features and the molecular diversities of DPMS.     

海平面上升对生态系统服务价值的影响及适应措施

海平面上升导致了海岸线后退、沿海侵蚀、风暴潮加强、生物栖息地改变、湿地变迁等,引起了近海域生态系统服务价值的变化。以美国Hillsborough County为研究区域,应用SLAMM模型,模拟了海平面从2010年—2100年上升1m的情景下各类用地面积的变化,利用效益转移法评价了生态系统服务价值的变化。结果表明:1旱地面积减少3037 hm~2,湿地面积增加3037hm~2,其中河口水域、滩涂、盐沼、定期洪水沼泽面积大幅度增加;季节性洪水沼泽、潮汐淡水沼泽、河流潮汐、沙滩面积大幅度减少,其它类型湿地受影响较小。2湿地总面积的增加反而导致了生态系统服务价值总量的降低,从2010年的61672万美元降低到2100年的61548万美元,这是由于净水湿地和沙滩大面积损失引起的;其中艺术娱乐、水调节、气候调节、文化精神价值将下降,水供应、栖息地保护、干扰调节和废物处理价值将上升。3防护、适应、后退3种措施分别降低、不影响、提升了生态系统服务价值,应根据其适用范围综合应用以制定应对海平面上升的适应性措施。     

基于林龄的滨海盐碱地杨树刺槐混交林土壤理化性质及草本植物多样性动态

以黄河三角洲滨海盐碱地不同林龄的杨树(Populus×Euramercana‘Neva’)、刺槐(Robinia pseucdoacacia)人工混交林及相应的纯林为研究对象,采用空间代时间的方法,研究了其不同林龄人工林林下植被特征与土壤理化性质变化。结果表明:(1)白茅(Imperata cylindrica)、猪毛菜(Salsola collina)、刺儿菜(Cirsium setosum)是滨海盐碱地人工林林下群落的优势种。(2)随着林分年龄(3年、7年、18年)的增加,杨树刺槐混交林及相应纯林Patrick指数及Margalef指数、Shannon-Wiener指数、Simpson指数及Pielou指数均表现出低-高-低的变化趋势,7年生时丰富度、草本多样性、均匀度最高。(3)杨树刺槐混交提高了林下草本物种组成及重要值,就纯林而言,刺槐林下植被物种多样性显著高于同龄杨树。(4)随着林龄的增大,杨树刺槐混交林及其纯林林下土壤p H值逐渐降低,杨树刺槐纯林土壤含盐量先增后减,杨树×刺槐混交林土壤含盐量依次递减;杨树刺槐混交林及其纯林林下土壤全磷含量呈先增加后减小趋势;杨树纯林及杨树×刺槐混交林全氮含量呈先减小后增加的趋势,刺槐纯林林下土壤全氮量呈连续增长的趋势;林龄间土壤p H、含盐量、全氮及全磷含量差异显著。(5)草本植物多样性相关指数与土壤含盐量呈极显著负相关,与土壤全磷量呈极显著正相关。     

黄海山东海域春、秋季鱼类群落多样性

根据2006年春季(5月)和秋季(10月)黄海山东海域疏目变水层双拖网调查数据,作者对该海域鱼类群落多样性、空间分布及长度谱特征进行了研究。调查共捕获鱼类61种,其中鲈形目种类最多(27种),其次为鲱形目(8种)和鲉形目(7种);按摄食类型可分为浮游动物食性、浮游动物/底栖动物食性、底栖动物食性、底栖动物/游泳动物食性、游泳动物食性、杂食性、碎屑食性7个类群,其中浮游动物食性占渔获物重量组成的82.5%。春、秋季鱼类群落均主要由小型种类占优势,春季优势种有3种,为玉筋鱼(Ammodytes personatus)、鳀(Engraulis japonicus)和方氏锦鳚(Pholis fangi),秋季优势种有5种,分别是鳀、青鳞小沙丁鱼(Sardinella zunasi)、玉筋鱼、小黄鱼(Larimichthys polyactis)和赤鼻棱鳀(Thryssa kammalensis)。丰富度指数(D)、Shannon-Wiener多样性指数(H’)、均匀度指数(J’)的空间异质性明显,表现出近岸海域(尤其是青岛-乳山南部近岸海域)较高,深水区较低的态势,但多样性指数的季节间差异不显著。资源量与Shannon-Wiener多样性指数(H’)、均匀度指数(J’)、功能团多样性指数(FD)均呈负相关关系,与种类丰富度指数(D)则无相关关系。春、秋季鱼类群落长度谱曲线斜率均小于-0.2,而截距均大于10,与该海域1985年及2000年的研究结果相比,长度谱斜率显著降低、截距显著增加,这与小型个体种类占优势密切相关。与历史资料相比,黄海山东海域鱼类群落多样性下降,小型个体比例上升,资源呈衰退趋势。     

中国机动车总颗粒物排放时空演变特征及驱动因子

机动车尾气颗粒物排放已成为城市空气污染的主要来源,严重影响环境空气质量和人体健康。研究基于2011-2015年中国市级机动车总颗粒物（total particulate matter,TPM）排放量数据,运用空间分析方法从存量和增量角度探究中国机动车TPM排放的时空演变特征,进而利用地理探测器模型定量评价其主要驱动因素的影响强度。结果表明：中国机动车TPM排放量呈逐年降低趋势,省会和直辖市是中国机动车TPM排放量减少的主要贡献者。省会和直辖市的机动车TPM排放量和减排量最大,其各年均值均超全国市级年均值的2倍。中国机动车TPM排放量和减排量均呈东部沿海至西部内陆地区递减趋势,且呈"低排放,高增加;高排放、高减少"的空间分布特征。"低排放,高增加"区域主要集中在中西部地区,特别是西南各省市;"高排放、高减少"区域为以京津冀为核心的泛华北平原地区。机动车TPM排放量的空间集聚性下降,空间随机分布趋势加强,热点区破碎化的空间分布格局日趋显著。地理探测分析表明,机动车TPM排放量受机动车数量的驱动作用最强,其次是机动车的使用强度,受自然环境条件的驱动作用最弱。年平均气温和海拔等自然因素主要通过与人类活动的共同作用显著增强对机动车TPM排放量的解释力。探究机动车颗粒物排放的时空异质特征和驱动因素,对提高中国机动车尾气治理的精准性具有重大意义。     

多毛类底栖动物在莱州湾生态环境评价中的应用

多毛类底栖动物常作为环境扰动的指示生物.对2011年莱州湾20个站位的多毛类底栖动物群落生态特征及其与沉积物理化特征的相互关系进行了研究,以期应用于莱州湾生态环境质量状况评价.结果表明,莱州湾多毛类共60种,4季平均生物密度为548.7个/m2,平均生物量为2.79 g/m2,机会种种类数量占优势种种类37.5％.种类多样性较高站位主要集中在湾东部,较低站位集中在湾西部和南部.ABC曲线表明多毛类动物处于中等扰动状态.BOPA指数显示,春季,站位S15和S16为中度污染,夏季,S14站位为重度污染.BIO-ENV分析结果显示,5个最佳环境变量组合相关系数ρs＞0.8,表明沉积物理化性质对多毛类动物群落结构的影响是显著的,沉积物粒度是最主要环境因子.因此,可通过多毛类种类、数量组成的时空变化特征及其与环境因子的生态关系,评价莱州湾生态环境质量状况,为环境污染控制和海洋生态环境保护提供基础信息和科学依据.     

系统型硬化症尿液lncRNA-mRNA差异表达基因筛选及生物信息学分析

为探讨系统性硬化症(SSc)患者尿液样本中的长链非编码RNA(lncRNA)、信使RNA(mRNA)的表达谱和生物学功能。选取6名SSc患者和3名健康对照者(HC),采集样本为中段晨尿,应用mRNA和lncRNA微阵列检测总RNA表达变异,SSc组与HC组相比。检测尿液lncRNA和mRNA表达,Gene ontology (GO)分析Kyoto Encyclopedia of Genes and Genomes (KEGG)信号通路分析差异表达的lncRNA功能分布;STRING在线网站和Cytoscape软件网络应用分析构建蛋白质相互作用网络(PPI)并筛选出核心基因(Hub Gene)。结果发现：与HC相比,SSc患者尿液中共有645个(上调546,下调99)mRNA和1 888个(上调1 647,下调241)lncRNA差异表达(Fold Change绝对值≥2,且P≤0.05)。KEGG通路结果显示富集TGF-β信号通路、氧化磷酸化、磷酸戊糖通路。SSc的GO分析显示与转录调控、DNA去甲基化、白介素6反应等相关;PPI网络分析表明主要富集在氧化磷酸化、细胞凋亡、自噬途径通路...