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121.
Nara T Hashimoto M Hirawake H Liao CW Fukai Y Suzuki S Tsubouchi A Morales J Takamiya S Fujimura T Taka H Mineki R Fan CK Inaoka DK Inoue M Tanaka A Harada S Kita K Aoki T 《Biochemical and biophysical research communications》2012,418(1):140-143
The first 3 reaction steps of the de novo pyrimidine biosynthetic pathway are catalyzed by carbamoyl-phosphate synthetase II (CPSII), aspartate transcarbamoylase (ATC), and dihydroorotase (DHO), respectively. In eukaryotes, these enzymes are structurally classified into 2 types: (1) a CPSII-DHO-ATC fusion enzyme (CAD) found in animals, fungi, and amoebozoa, and (2) stand-alone enzymes found in plants and the protist groups. In the present study, we demonstrate direct intermolecular interactions between CPSII, ATC, and DHO of the parasitic protist Trypanosoma cruzi, which is the causative agent of Chagas disease. The 3 enzymes were expressed in a bacterial expression system and their interactions were examined. Immunoprecipitation using an antibody specific for each enzyme coupled with Western blotting-based detection using antibodies for the counterpart enzymes showed co-precipitation of all 3 enzymes. From an evolutionary viewpoint, the formation of a functional tri-enzyme complex may have preceded-and led to-gene fusion to produce the CAD protein. This is the first report to demonstrate the structural basis of these 3 enzymes as a model of CAD. Moreover, in conjunction with the essentiality of de novo pyrimidine biosynthesis in the parasite, our findings provide a rationale for new strategies for developing drugs for Chagas disease, which target the intermolecular interactions of these 3 enzymes. 相似文献
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123.
T. WORTHINGTON P. S. KEMP P. E. OSBORNE A. DILLEN J. COECK M. BUNZEL‐DRÜKE M. NAURA J. GREGORY K. EASTON 《Freshwater Biology》2012,57(3):602-611
1. Availability of suitable habitat is a prerequisite for species reintroduction success, and to ensure population persistence, investigations of a species’ habitat utilisation throughout its life history should be conducted as part of a feasibility study. 2. Habitat utilisation models for burbot, Lota lota, developed using data from field studies conducted in France and Germany and information from the literature were used to assess the feasibility of reintroducing burbot into rivers of its former native range in eastern England. 3. Per cent tree roots, aquatic vegetation and flow types were important predictors of adult burbot abundance. Furthermore, the habitat utilisation models were supplemented with information from the literature, which suggested that off‐channel habitat such as wetlands and backwaters is important for spawning and nursery stages. 4. An assessment of the habitat availability in the rivers of the burbot’s former native range using variables related to spawning and nursery and adult life stages showed that although adult habitat was widely distributed, the availability of spawning and nursery habitat was less abundant, potentially limiting successful reestablishment. 5. Potential suitable habitat was concentrated in the central and southern areas of the species’ former English distribution. Overall, rivers of the burbot’s former range potentially afford suitable habitat to sustain a reintroduced population. However, sites should be preferentially selected on the basis of having appropriate spawning and nursery areas. 相似文献
124.
Suenaga K Lee KY Nakamori M Tatsumi Y Takahashi MP Fujimura H Jinnai K Yoshikawa H Du H Ares M Swanson MS Kimura T 《PloS one》2012,7(3):e33218
Myotonic dystrophy type 1 (DM1) is a multi-systemic disorder caused by a CTG trinucleotide repeat expansion (CTG(exp)) in the DMPK gene. In skeletal muscle, nuclear sequestration of the alternative splicing factor muscleblind-like 1 (MBNL1) explains the majority of the alternative splicing defects observed in the HSA(LR) transgenic mouse model which expresses a pathogenic range CTG(exp). In the present study, we addressed the possibility that MBNL1 sequestration by CUG(exp) RNA also contributes to splicing defects in the mammalian brain. We examined RNA from the brains of homozygous Mbnl1(ΔE3/ΔE3) knockout mice using splicing-sensitive microarrays. We used RT-PCR to validate a subset of alternative cassette exons identified by microarray analysis with brain tissues from Mbnl1(ΔE3/ΔE3) knockout mice and post-mortem DM1 patients. Surprisingly, splicing-sensitive microarray analysis of Mbnl1(ΔE3/ΔE3) brains yielded only 14 candidates for mis-spliced exons. While we confirmed that several of these splicing events are perturbed in both Mbnl1 knockout and DM1 brains, the extent of splicing mis-regulation in the mouse model was significantly less than observed in DM1. Additionally, several alternative exons, including Grin1 exon 4, App exon 7 and Mapt exons 3 and 9, which have previously been reported to be aberrantly spliced in human DM1 brain, were spliced normally in the Mbnl1 knockout brain. The sequestration of MBNL1 by CUG(exp) RNA results in some of the aberrant splicing events in the DM1 brain. However, we conclude that other factors, possibly other MBNL proteins, likely contribute to splicing mis-regulation in the DM1 brain. 相似文献
125.
Fujimura Yoshiyasu Fujita Hiroko Takada Masayuki Inoue Takashi 《Landscape and Ecological Engineering》2012,8(2):215-221
We examined the relationship between fluctuation patterns of groundwater levels (WL) and the distribution dynamics of the vascular plant Sasa palmata, in Sarobetsu Mire, northern Japan. WLs were recorded at 30 stations: 11 Sphagnum lawns, which is the original vegetation type in the area, nine Sasa communities, and ten boundary stations between those two areas. The ten boundary stations were composed of five vegetation change sites and five stable sites, categorized using maps of Sasa distribution in 1977 and 2000. The results showed that Sphagnum lawns and Sasa communities significantly differed in the average, minimum, and fluctuation range of WLs, and increases in WL after rain events. The differences between change sites and stable sites were not significant in the average WL, but were significant in the minimum and fluctuation ranges of WL and the increase in WL after rain events. These results indicate that the distribution dynamics of Sasa communities would be regulated by groundwater flow that was affected by drainage and inflow condition rather than merely groundwater drawdown as suggested in previous studies. The WL regimes at two Sphagnum lawns were similar to those at change sites, suggesting that Sasa may expand into these two stations more readily than the other Sphagnum lawn stations. 相似文献
126.
To date, minerals of interest have been analyzed individually to understand mineral dynamics and metabolism. Our recent development of metallomic analyses enabled us to evaluate minerals in an unbiased and global manner. Here, we evaluated the effects of ingestion of excess zinc to plasma and tissue concentrations of minerals in growing rats. A total of 26 minerals were simultaneously evaluated by metallomic analyses using inductively coupled plasma-mass spectrometry (ICP-MS) in semi-quantification mode; the concentrations of several minerals exhibited consistent changes in response to the concentrations of dietary zinc. Manganese concentrations in plasma and femur increased, while concentrations in the liver and pancreas decreased with increasing dietary zinc concentrations. Because the interaction between zinc and manganese is not known, we further focused our analysis on liver manganese. Quantitative analyses also indicated that the hepatic concentration of manganese decreased in response to the ingestion of diets containing excess zinc, a result that is partly explained by the decreased expression of hepatic Zip8, a manganese transporter. The present study reveals mineral interaction by using metallomic analyses and proposes a possible mechanism that underlies this novel interaction. 相似文献
127.
高维、小样本数据的特征选择方法在蛋白质质谱数据处理分析领域有着广泛应用。本文针对蛋白质质谱特征选择问题,结合稀疏表示这一新理论框架,提出了一种基于稀疏表示的特征选择算法(sparse representation based feature selection,SRFS)。该方法将稀疏表示分类的结果作为评定某一个特征子空间特征相对重要性的度量,然后通过对大量随机采样子空间计算结果的统计,得到特征空间中每个特征的排序,并进一步分析提炼出与肿瘤疾病相关的若干谱峰。通过在卵巢癌公共数据集OC-WCX2a和浙江省肿瘤医院乳腺癌数据集BC-WCX2a上的实验结果表明,SRFS算法可以有效应用于本文所使用的SELDI-TOF蛋白质质谱数据的分析。 相似文献
128.
YANG Zhang-ping CAI Shao-li KE Chong-rong HUANG Ping LIU Xiao-lin YU Qing-mei ZHUANG Xiu-hong CHEN Lin LIN Guo-yan HUANG Jian-zhong 《工业微生物》2012,42(3)
通过对木聚糖酶高产菌株EIM-30基于形态学和18S rDNA序列的系统发育进化分析,鉴定为里氏木霉Trichoderma reesei.在单因子实验确定EIM-30产木聚糖酶的最适碳源和氮源的基础上,通过Plackett-Burman实验对影响其产酶的相关因素进行评估并筛选出3个显著效应因子,然后应用最陡爬坡实验和响应面分析确定最适产酶培养基配方为:酵母浸膏1.50%,蛋白胨1.00%,NaCl 0.50%,PPG-2000 0.10%,MgSO4 1.20%,CaCl2 0.18%,(NH4)2 SO4 0.45%,甘油4.18%,乳糖3.05%,K2HPO41.59%.优化后Trichoderma reesei EIM-30的液体发酵产木聚糖酶的活力可达9.857×105 U/mL,较优化前提高1.98倍. 相似文献
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130.
采用冻干浓缩、(NH4)2S04盐析、HiTrapphenyl(FF)疏水层析和QSepharose FastFlow离子交换层析对灵芝EIM-40发酵液进行分离纯化,获得纯化漆酶,纯化倍数为14.6,回收率为5.3%。SDS-PAGE银染的结果为单一条带,相对分子质量约为6.53×104。以愈创木酚和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)为催化底物进行酶学性质研究,最适pH分别为4.8和4.5,最适温度分别为55和50℃,2种底物在pH4.0。5.0范围内,温度低于50℃时,酶的稳定性都很好。以愈创木酚为底物,Km=645.0umol/L;以ABTS为底物,Km=22.2txmol/L。Cu2+对该酶起激活作用,Fe2+、Ca2+、Ba2+则完全抑制酶的活性。 相似文献