Molecular characterization of a thermostable aldehyde dehydrogenase (ALDH) from the hyperthermophilic archaeon Sulfolobus tokodaii strain 7

Aldehyde dehydrogenase (ALDH) is a widely distributed enzyme in nature. Although many ALDHs have been reported until now, the detailed enzymatic properties of ALDH from Archaea remain elusive. Herein, we describe the characterization of an ALDH from the hyperthermophilic archaeon Sulfolobus tokodaii. The enzyme (stALDH) could utilize various aldehydes as substrates, and maximal activity was found with acetaldehyde and the coenzyme NAD. The optimal temperature and pH were 80 °C and 8, respectively, and high thermostability was found with the half-life at 90 °C to be 4 h. The enzyme was considerably resistant to nitroglycerin (GTN) inhibition, which could be restored by reducing agent DTT or (±)-??-lipoic acid. Coenzyme NAD or NADP could regulate the enzymatic thermostability, as well as the esterase activity. Molecular modeling suggested that the enzyme harbored similar structural arrangement with its eukaryotic and bacterial counterparts. Sequence alignment showed the conserved catalytic residues E240 and C274 and cofactor interactive sites N142, K165, I168 and E370, the function of which were verified by site-directed mutagenesis analysis. This is the most thermostable ALDH reported until now and the unique property of this enzyme is potentially beneficial in the fields of biotechnology and biomedicine.     

Tethered Spectroscopic Probes Estimate Dynamic Distances with Subnanometer Resolution in Voltage-Dependent Potassium Channels

Measurements of inter- and intramolecular distances are important for monitoring structural changes and understanding protein interaction networks. Fluorescence resonance energy transfer and functionalized chemical spacers are the two predominantly used strategies to map short-range distances in living cells. Here, we describe the development of a hybrid approach that combines the key advantages of spectroscopic and chemical methods to estimate dynamic distance information from labeled proteins. Bifunctional spectroscopic probes were designed to make use of adaptable-anchor and length-varied spacers to estimate molecular distances by exploiting short-range collisional electron transfer. The spacers were calibrated using labeled polyproline peptides of defined lengths and validated by molecular simulations. This approach was extended to estimate distance restraints that enable us to evaluate the resting-state model of the Shaker potassium channel.     

Diffusive greenhouse gases fluxes from the surface of the Three Gorges Reservoir: Study at a site in Fuling

To investigate the water-air diffusive greenhouse gases (GHGs) fluxes from the Three Gorges Reservoir (TGR), a field experiment on carbon dioxide (CO₂), methane (CH₄) and nitrous oxide (N₂O) fluxes from water surface was carried out from March 2011 to August 2012 by floating static chamber method. The results showed that CO₂ was released to the atmosphere all the time and was less in autumn than in other seasons (P < 0.05). CH₄ was also released to the atmosphere throughout the year but more in summer than other three seasons (P < 0.05). N₂O flux was higher in autumn than other seasons (P < 0.05), and N₂O was absorbed from the atmosphere mainly in summer. Moreover, correlation analysis illustrated that CO₂ flux had significantly negative correlation with wind velocity (P < 0.05), whereas positive correlation with pH (P < 0.01) had been found. There was no significant correlation between CH₄ (or N₂O) flux and the measured environmental variables respectively (P > 0.05). Additionally, the annual fluxes of CO₂, CH₄ and N₂O were 140.45 ± 12.57 mg CO₂·m^?2 h^?1, 1.35 ± 0.14 mg CH₄·m^?2 h^?1 and 34.34 ± 11.64 μg N₂O·m^?2 h^?1, respectively. When compared to other reservoirs worldwide, the CO₂ and N₂O fluxes from TGR were higher than those from boreal and temperate reservoirs, but much lower than those from tropical reservoirs. CH₄ flux was lower than those from boreal, temperate and most tropical reservoirs. In our study, the surface area of the TGR emitted 1.42 × 10⁶ t CO₂, 1.19 × 10⁴ t CH₄ and 589.93 t N₂O in a year. The total GWP was 17.68 t CO₂-eq ha^?1 yr^?1, of which CO₂ flux was dominant (74.38%). Therefore, CO₂ was the main contributor of GHGs fluxes in our study and thus future researches should focus on how to reduce CO₂ fluxes from the surface of the TGR. TGR has a considerable contribution to regional GHG emissions.     

水牛精子蛋白质组双向电泳体系的建立和优化

建立和优化一种适合水牛精子蛋白质组学研究的双向电泳技术。以水牛精子为研究对象,比较两种不同配方的裂解液,以及不同上样量对其2-DE图谱质量的影响。结果显示,以7 mol/L尿素、2 mol/L硫脲、4%CHAPS、1%DTT、0.5%Cocktail of protease inhibitors为裂解液,24 cm胶条上样量200μg时,可获得较好的精子总蛋白质2-DE图谱。运用ImageMaster 2-Dplatinum分析软件检测出约500个蛋白质点,蛋白质大部分分布在等电点5-7之间,分子量范围约40-90 kD。     

甲状腺髓样癌遗传学研究进展

甲状腺髓样癌(medullary thyroid carcinoma,MTC)是起源于甲状腺C细胞或滤泡旁细胞的恶性肿瘤,分遗传型髓样癌和散发型髓样癌两种.MTC主要由RET原癌基因突变引起,对患者进行基因测序分析能在基因水平诊断MTC,从而为患者早期行预防性手术治疗提供依据.从甲状腺髓样癌临床分型、分子遗传基础及动物模型不同层面进行综述,有利于进一步了解疾病致病机制和开展药物实验性治疗研究.     

林分耗水的尺度扩展研究进展

蒸散耗水是森林生态系统水分循环过程的主要构成,是现代生态水文研究的重点和难点。直接测定和尺度上推提供了两种获取蒸散量的技术手段。受限于复杂的冠层结构、非均质的下垫面和迥异的环境条件,直接测定难以准确获取林分水平的耗水信息,因此有必要讨论单木与冠层结构、环境因子之间的耦合关系,利用时空尺度扩展得到合理的耗水量。本文综述了理论基础相对牢固的3种尺度扩展技术:基于生物计量参数、遥感影像和水文模型的尺度扩展,剖析各模型的控制因子,探讨模型的适用性和优缺点,对尺度扩展技术的发展做了展望。