Initial Molecular Recognition Steps of McjA Precursor during Microcin J25 Lasso Peptide Maturation

Microcin J25 (MccJ25) has emerged as an excellent model to understand the maturation of ribosomal precursor peptides into the entangled lasso fold. MccJ25 biosynthesis relies on the post‐translational modification of the precursor McjA by the ATP‐dependent protease McjB and the lactam synthetase McjC. Here, using NMR spectroscopy, we showed that McjA is an intrinsically disordered protein without detectable conformational preference, which emphasizes the active role of the maturation machinery on the three‐dimensional folding of MccJ25. We further showed that the N‐terminal region of the leader peptide is involved in interaction with both maturation enzymes and identified a predominant interaction of V43–S55 in the core McjA sequence with McjC. Moreover, we demonstrated that residues K23–Q34 in the N‐terminal McjA leader peptide tend to adopt a helical conformation in the presence of membrane mimics, implying a role in directing McjA to the membrane in the vicinity of the lasso synthetase/export machinery. These data provide valuable insights into the initial molecular recognition steps in the MccJ25 maturation process.     

On the association between poly(vinyl alcohol) and sodium dodecyl sulfate and its effect on liquid–liquid interfacial tension: A mathematical model

Association between poly(vinyl alcohol-co-vinyl acetate) copolymer (PVA) and sodium dodecyl sulfate (SDS) was studied experimentally and theoretically. It was found that, for the ethyl acetate-aqueous phase interface in which PVA was previously adsorbed, the interfacial tension (γ) increases abruptly to a maximum and then exhibits a relatively mild decay with the addition of SDS to the aqueous phase. The theoretical results indicate that vinyl acetate (VAc) segments determine γ. However, for relatively low concentrations of SDS (C_SDS), this latter plays a major role because through its association with the VAc segments it modulates the extent to which PVA is adsorbed at the interface, indirectly determining the value of γ. As C_SDS approaches to the CMC value for SDS, its influence on γ decreases because SDS tends to self-assembly rather than associates with VAc. These model predictions are consistent with experimental findings reported in the literature.     

Efficient loading and entrapment of tamoxifen in human serum albumin based nanoparticulate delivery system by a modified desolvation technique

Herein, the poorly water-soluble drug, Tamoxifen (Tmx), was loaded in the amphipathic matrix of human serum albumin (HSA) nanoparticles by a modified desolvation method. In order to enhance the drug loading (DL) and drug entrapment efficiency (DEE) (<2% and 10%, respectively), ultrasonication of Tmx-HSA mixture was performed prior to desolvation process. Tmx loading and entrapment efficiency were optimized by employment of the response surface methodology (RSM)-central composite design (CCD) of experiments. Under the optimum conditions of 1.59 mg Tmx/ml concentration, 7.76 pH and 5 h incubation of HSA-Tmx, the DL of 6.7% and DEE of 74% are achievable. Particles with the average size of 195 nm, zeta potential of −21 mV and polydispersity index of 0.09 were produced under these conditions. A more sustained Tmx release behavior was observed from polyethylene glycol (PEG) conjugated nanoparticles in comparison to the non-PEGylated ones. The short-term stability investigation showed no alteration in physicochemical properties of nanoparticles at 4 and 37 °C, but small increase in nanoparticles size was observed after three months of storage at room temperature. This is the first report for efficient production of a Tmx delivery system based on HSA nanoparticles.     

Newly designed polyacrylamide/dextran gels for electrophoresis protein separation: synthesis and characterization

Newly designed gels for electrophoresis protein separation were synthesized from acrylamide, N,N′‐methylenebis (acrylamide) and dextran mixtures. Radical polymerization was initiated by ammonium persulfate and N,N,N′,N′‐tetramethylethylenediamine. The time dependence of absorbance during polymerization was monitored using UV‐visible spectroscopy. The exothermic polymerization process exhibited a sharp rise of temperature reminiscent of the Trommsdorff effect. The swelling kinetics of the synthesized gels was examined in deionized water and buffer solutions. One of the challenges was to find an alternative to commercial products, sold as mixtures with no detailed chemical contents, commonly used in sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS‐PAGE) for protein separation. For this reason, a systematic comparison was made of the properties of one of the most commonly used commercial gels, Duracryl^? from Genomics Solution Inc., and those of the synthesized polyacrylamide/dextran gels. Copyright © 2011 Society of Chemical Industry     

Purification and characterization of parvalbumins from silver carp (Hypophthalmichthy molitrix)

BACKGROUND: As the largest producer and consumer of freshwater fish in the world, many people suffer from allergy for consuming freshwater fish in China. However, the allergen profiles of freshwater fish are rarely known. RESULTS: Parvalbumins (PVs) from the white muscle of silver carp (Hypophthalmichthy molitrix) were purified by ammonium sulfate fractionation and column chromatography including DEAE‐Sepharose and Superdex 75. Three PV isoforms—PV‐I, PV‐II, and PV‐III—were obtained and their molecular masses as estimated by tricine–sodium dodecyl sulfate–polyacrylamide gel electrophoresis were 12, 11, and 14 kDa, respectively. All the PVs could be detected by anti‐frog PV monoclonal antibody. PV‐I and PV‐II were quite possibly glycoproteins, while PV‐III was not glycosylated, as analyzed by periodic acid–Schiff (PAS) staining. Thermal stability revealed that PV‐I and PV‐II easily formed polymers, while these proteins were stable in a pH range of 4.0–10.0. A PV gene encoding 110 amino acid residues was cloned and it revealed high identity with PVs from other species of fish. CONCLUSION: Three isotypes of PV were purified to homogeneity and one distinct PV gene was cloned in silver carp white muscle. Copyright © 2010 Society of Chemical Industry     

Purification and structural characterisation of lipid transfer protein from red wine and grapes

Lipid transfer proteins (LTP) play a major role in plant defence and are of particular interest due to their known ability to cause allergic reactions. These proteins are expressed in grapes and also remain detectable after vinification, especially in red wine. However, it remains unknown whether the protein undergoes any changes during the vinification process. Here, we present a purification method for LTPs from Dornfelder grapes and wine. By liquid-chromatography–mass spectroscopy (LC–MS/MS) we identified LTPs from two different species (Vitis vinifera and Vitis aestivalis). Additionally, the purified LTPs were characterised using spectrometric methods, confirming their high purity and structural stability during vinification. We conclude that LTPs are resistant to the alcohol content (13.5 vol%), acidic milieu of wine and other ingredients present during the vinification process, indicating that the allergenic potential of grape LTP is not diminished by the vinification process.     

TETRA电路模式控制实体信令软件实现

TETRA作为全球性数字集群标准,其技术和市场都在迅速发展。本文详细分析了TETRA移动台电路模式控制实体(CMCE-MS)协议,并给出其软件设计与实现的详细方案。     

珊瑚状分级微纳米结构聚苯胺的制备、表征及其气敏性能研究

采用简单的阴离子表面活性剂诱导原位化学聚合法,以过硫酸铵(APS)为氧化剂,在十二烷基硫酸钠(SDS)的盐酸溶液中制得了珊瑚状聚苯胺( PANI)微纳米材料.对所制备的PANI分别采用傅里叶变换红外光谱(FT-IR)、场发射扫描电镜(FE-SEM)进行表征.实验结果表明加入SDS后,可以得到一种珊瑚状分级结构PANI微...     

高光谱空间降采样独立成分特征分离

提出一种空间降采样独立成分特征分离方法,用于缩短独立成分分析(ICA)法在高光谱图像特征分离时的运算时间。该方法通过对高光谱图像的二维像素空间进行网格划分得到较小的窗口;基于光谱相似性测度法度量每个窗口中中心像素与周围像素的距离。然后舍弃这段距离小于阈值的周围像素,而将大于阈值的周围像素和中心像素作为样本量进行FastICA,获取投影矩阵变换原始数据,得到特征分离的ICA成分。对比了传统ICA与空间降采样(SDS)ICA(SDS_ICA)的性能,研究了降采样阈值参数、降采样窗口参数及初始投影矩阵对SDS_ICA特征分离性能及运行时间的影响。实验结果表明:应用SDS_ICA时,仅设置适中的阈值和不敏感的窗口大小参数,就能保持与传统ICA相近的特征分离性能,运行时间减少了30%以上。该方法适合应用于高光谱准实时特征提取、数据降维及目标探测等领域。