亚甲蓝双分子聚集体的解离平衡

本通过光主地研究了作为新的光敏剂－亚甲蓝（Ｍｅｔｈｙｌｎｅ Ｂｌｕｅ，ＭＢ）在水溶液中的溶液状态，以及其在不同温度时双分子聚集体的解离平衡，通过差示光谱监测了亚甲蓝在水溶液中，其双分子二聚体在温度３０－６０℃范围内的构成方式的变化：其单分子倾角９α）为４８．３－４６．１°，双分子面间距离（ｒ）为０．５７６－０．５８０ｎｍ，监测了亚甲蓝在水溶液中其单最大吸收波长吸收值随温度的变化，算得其二聚体的解     

树枝聚醚改性聚丙烯酰胺和阴离子表面活性剂的缔合行为

采用粘度法、荧光探针技术和^1H NMR驰豫和自扩散方法，研究了树枝聚醚疏 水改性丙烯酰胺共聚物(PDAM)和十二烷基硫酸钠(SDS)在水溶液中的相互作用．这 种共聚物含有少量的树枝聚醚，具有疏水性，容易和SDS发生相互作用，在表面活 性剂浓度远低于临界胶束浓度(cmc)的情况下，生成混合胶束状聚集体．它们的缔 合行为和溶液性质明显地取决于表面活性剂的浓度，随着聚合物溶液中加入SDS， 溶液粘度发生急剧变化，并在较低的表面活性剂浓度处出现很大的最高点．荧光和 ^1H NMR测定结果表明，这是由于在不同SDS浓度范围内，PDAM/SDS形成的聚集体结 构不同的缘故．     

甲醇-水、乙醇-水体系聚集态的共振散射光谱

自从Ｐａｓｔｅｒｎａｃｋ[1,2]使用普通荧光分光光度计建立共振光散射(ＲＬＳ)技术以来,人们用该技术建立了十分灵敏的蛋白质和核酸的分析方法[3,5]。同时该技术在研究化合物在溶液中的聚集态方面也有了较为广泛的应用,一般认为分子聚集体的形成是引起ＲＬＳ增强的主要原因[6]。已知甲醇、乙醇与水互溶形成均匀的溶液体系,那么在这些溶液体系中是否也存在某种形式的分子聚集体呢?为此本文研究了不同浓度的甲醇水溶液和乙醇水溶液的ＲＬＳ光谱。1　实验部分1 1　试剂和仪器甲醇(上海建鑫化工试剂厂,分析纯,含量%≥99 5);乙醇(安徽特酒总…     

D-，L-苯丙氨酸诱导非手性菁染料的手性组装

超分子手性与分子自组装是生命体中非常重要和有趣的现象．报道了D,L-苯丙氨酸等氨基酸在氯化钠溶液中通过非共价键相互作用诱导非手性菁染料（Pseudoisocyanine,PIC）J-聚集体超分子手性的形成．实验结果表明,诱导的手性菁染料PIC聚集体发色团在π-π^＊跃迁区域产生了特征的镜像圆二色性,其圆二色信号和强度强烈地依赖于氨基酸的绝对构型、浓度、侧链基团和溶液温度．原子力显微镜照片清楚地表明,^聚集体由相互交联的纳米纤维组成,诱导的圆二色性可能来源于纤维状聚集体的宏观螺旋排列．     

分子动力学模拟研究荧光分子芘在磺基甜菜碱胶束中的增溶

采用分子动力学模拟研究了荧光分子芘在磺基甜菜碱两性表面活性剂聚集体中的增溶现象．结果表明,芘分子自发地自溶液中增溶进入胶束疏水内核的栅栏层区域．当胶束溶液中芘分子的局部浓度增大时,两个芘分子可以同时增溶进胶束的栅栏层区域,此时两个芘分子形成π-π共轭堆积的激发态络合物．但是由于荧光分子之间的弱兀．兀相互作用,激发态络合物在胶束中是不稳定的,表现为两个芘分子的多次结合和分离．模拟表明,分子动力学方法可以在分子水平上研究荧光探针分子在表面活性剂胶束中的增溶位点,解释荧光分子在胶束中的动力学现象．     

分子间相互作用对蛋白质晶体生长的影响

采用原子力显微镜对溶菌酶和刀豆蛋白A的分子间相互作用力的情况进行了研究, 并用动态光散射研究了此二种分子间相互作用力有较大差异的蛋白质在晶体生长条件和非生长条件下, 溶液中的聚集体的状态(大小和分散度)随浓度和温度的变化情况. 实验结果表明, 范德华力强的刀豆蛋白A在成核前, 溶液中的聚集体不能很快转变为生长基元, 导致晶体生长时间长; 而范德华力弱的溶菌酶, 溶液中的聚集体可以很快转变成生长基元, 晶体生长时间也较短.     

含Gemini表面活性剂结构单元的新型两亲聚电解质的合成及聚集行为

分别将Gemini型单体1,3-双(二甲基十四烷基溴化铵)-2-丙烯酰氧基丙烷(14G)或1,3-双(二甲基十六烷基溴化铵)-2-丙烯酰氧基丙烷(16G)与丙烯酰氧乙基三甲基氯化铵(DAC, D)共聚合反应, 合成了新型含Gemini表面活性剂结构单元的两亲性阳离子聚电解质(D14G和D16G). 采用稳态荧光、电导、动态光散射及透射电镜等手段研究了这些聚电解质在水溶液中的聚集行为. 结果表明, 临界聚集浓度(CAC)随着Gemini型表面活性剂单元含量的增加而减小, 同时随着Gemini型表面活性剂单元中疏水碳链长度的增加而降低. 这些聚电解质在水溶液中同时存在分子内和分子间两种类型的聚集体, 而且碳链越长, 形成分子内聚集体的倾向越强. 随着Gemini表面活性剂单元含量的增加, D14G溶液中聚集体的流体动力学半径(Rh)也有所增大, 而D16G溶液中的聚集体的流体动力学半径(Rh) 却略有减小.     

光调控分子有序聚集体及其功能化研究进展

双亲分子在溶液中可以缔合形成胶束、囊泡、液晶、乳液等有序分子聚集体。在分子中引入功能性的基团,通过改变分子的结构、浓度或引入外部刺激,可以对有序分子聚集体的类型和性能进行调控。光作为一种绿色可控的清洁能源,是一种理想的外部刺激信号。在双亲分子中引入感光基团,可以通过光照调节有序聚集体的组装,并进一步实现功能性的调控。本文综述了近年来在光调控分子有序聚集体方面的研究及其在生物、传导、纳米材料制备中的应用。同时,对光调控的功能性有序分子聚集体未来的发展前景进行了展望。     

NaCl对液-液扩散法生长溶菌酶晶体的影响

用动态光散射法研究了不同浓度NaCl对液—液扩散法生长溶菌酶晶体的影响， 并测量了晶体生长前后体系的Zeta电势．结果表明，NaCl浓度较高时，在溶菌酶溶 液—凝胶界面处会发生液液分层现象，溶液中一直存在较大的聚集体，生长出的晶 体质量较差．而在合适的NaCl浓度下，随着溶液Zeta电势降低，溶液中溶菌酶的大 的聚集体发生解聚集，生长出的晶体质量较高．     

含芳环的可聚合阳离子型Hydrotrope的自缔合行为和助溶性能

采用尼罗红(Nile red)染料为探针,利用吸收和发射光谱研究了具有不同烷芳基结构的可聚合阳离子型Hydrotropes(PCHs)的溶液性质,结果表明其在水溶液中易发生自缔合行为并具有两个阈值浓度:分子开始自缔合的临界浓度(CAC)(10-1—10-2 mol/L),开始大幅度提高助溶作用的最小助溶浓度(MHC)(>10-1 mol/L),两阈值浓度均随化合物分子结构中的疏水基团尺寸的增大而减小.两亲化合物聚集体的微环境极性测定的结果显示,其极性比表面活性剂胶束高,并且随着化合物浓度的增大不断下降,表明形成的聚集体结构比较疏松,且其分子堆砌密度随聚集体的增大而逐步升高.此外,通过自由基聚合得到相应的两亲聚合物仍保留Hydrotrope原有的性能,并具有更显著的助溶作用.     

Trichobitacin — a new ribosome-inactivating protein I. The isolation,physicochemical and biological properties of trichobitacin

From the press-residue of the fresh root tuber of Trichosanthes kirilowii Maim (Cucurbitaceae), a new ribosome-inactivating protein (RIP), trichobitacin, was isolated. It has the activity of RNA N-glycosidase and can inhibit the growth of human placental trophoblastic cells. Its molecular weight is 27,228 Da (ES-MS) and pI 9.6. It is a single chain basic RIP. Its amino acid composition was determined. It is a new RIP. It consists of 0.7～0.9% galactose and may be a glycoprotein. Its N- and C-terminal amino acid is Asp and Ala, respectively. Its N-terminal preliminary amino acid sequence has been determined.     

Trichobitacin-a new ribosome-inactivating protein Ⅰ.The isolation,physicochemical and biological properties of trichobitacin

From the press-residue of the fresh root tuber of Trichosanthes kirilowii Maxim (Cu-curbitaceae),a new ribosome-inactivating protein (RIP),trichobitacin,was isolated.It has the activity of RNA N-glycosidase and can inhibit the growth of human placental trophoblastic cells.Its molecular weight is 27,228 Da (ES-MS) and pI 9.6.It is a single chain basic RIP.Its amino acid composition was determined.It is a new RIP.It consists of 0.7~0.9% galactose and may be a glycoprotein.Its A'-and C-terminal amino acid is Asp and Ala,respectively.Its N-terminal preliminary amino acid sequence has been determined.     

Trichosanthin's interfacial interactions with phospholipids: a monolayer study

Lipid monolayer at the air/water interface, as half a membrane, was used here to investigate the interaction between trichosanthin (TCS), a ribosome inactivating protein, and phospholipid membrane. First, the protein adsorption experiments showed that the negatively charged DPPG caused obvious enrichment of TCS beneath the monolayer, indicating electrostatic attraction between TCS and the negatively charged phospholipid. Second, when TCS was incorporated into the phospholipid monolayer, it could not be completely squeezed out until the monolayer collapsed. The results were demonstrated to be irrelative with the phospholipid headgroup, suggesting a strong hydrophobic force between TCS and phospholipid hydrocarbon chain was involved in the interaction. Third, the protein/membrane interaction was further studied with fluorescence microscope. The results showed that TCS could penetrate into both the condensed and the fluid phase of the DPPG monolayer under low pH condition and eventually resulted in a homogeneous phospholipid phase. The breakage of ordered packing of phospholipid by TCS may be responsible for this homogenizing effect.     

Radiation Damage and Racemic Protein Crystallography Reveal the Unique Structure of the GASA/Snakin Protein Superfamily

Proteins from the GASA/snakin superfamily are common in plant proteomes and have diverse functions, including hormonal crosstalk, development, and defense. One 63‐residue member of this family, snakin‐1, an antimicrobial protein from potatoes, has previously been chemically synthesized in a fully active form. Herein the 1.5 Å structure of snakin‐1, determined by a novel combination of racemic protein crystallization and radiation‐damage‐induced phasing (RIP), is reported. Racemic crystals of snakin‐1 and quasi‐racemic crystals incorporating an unnatural 4‐iodophenylalanine residue were prepared from chemically synthesized d ‐ and l ‐proteins. Breakage of the C?I bonds in the quasi‐racemic crystals facilitated structure determination by RIP. The crystal structure reveals a unique protein fold with six disulfide crosslinks, presenting a distinct electrostatic surface that may target the protein to microbial cell surfaces.     

天花粉蛋白的聚乙二醇定点修饰及其性质的初步研究

用聚乙二醇(PEG)定点修饰天花粉蛋白(TCS), 并比较了修饰前后TCS的生物活性、免疫原性及药代动力学等诸多性质. 选择TCS分子上可能的抗原决定簇位点KR173-174进行定点突变, 并将所构建的突变体TCS_KR173-174CG在大肠杆菌中表达及纯化. 通过该突变体第173位引入的半胱氨酸残基进行PEG定点修饰. 通过比较研究, 分析所构建的PEG修饰型TCS的DNA酶活性、致核糖体失活活性、免疫原性、急性毒性以及药代动力学性质, 研究结果表明, 所构建的突变型TCS(mTCS)的活性与野生型TCS(wTCS)活性几乎相当, 而免疫原性已显著降低. PEG修饰型TCS虽有活性下降, 但其免疫原性、急性毒性以及药代动力学性质得到显著提升. 表明通过基因工程及化学修饰方法改造TCS是可行的, 所构建的突变型及PEG修饰型TCS值得进一步研究.     

Cloning, overexpression, purification, and characterization of receptor-interacting protein 3 truncation inEscherichia coli

To facilitate structural studies of receptor-interacting protein 3 (RIP3), we developed a large-scale expression system of a glutathione-S-transferase (GST) fused with an 82 amino acid RIP3 protein inEscherichia coli. RIP3 truncation was subcloned into the pGEX-4T-1 vector and overexpressed in BL21(DE3)RIL cells. The soluble RIP3 protein was successfully purified to homogeneity using GST tag, an anion-exchange column, and gel filtration chromatography. The purity, identity, and conformation of the RIP3 protein were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, matrix-assisted laser desorption ionization mass spectrometry, circular dichroism, and fluorescence spectroscopic studies. RIP3 showed dominance of the α-helix structure and temperature-dependent conformational change.     

Dynamic mechanism for the autophosphorylation of CheA histidine kinase: molecular dynamics simulations

The two-component system (TCS) is an important signal transduction component for most bacteria. This signaling pathway is mediated by histidine kinases via autophosphorylation between P1 and P4 domains. Taking chemotaxis protein CheA as a model of TCS, the autophosphorylation mechanism of the TCS histidine kinases has been investigated in this study by using a computational approach integrated homology modeling, ligand-protein docking, protein-protein docking, and molecular dynamics (MD) simulations. Four nanosecond-scale MD simulations were performed on the free P4 domain, P4-ATP, P4-TNPATP, and P1-P4-ATP complexes, respectively. Upon its binding to the binding pocket of P4 with a folded conformation, ATP gradually extends to an open state with help from a water molecule. Meanwhile, ATP forms two hydrogen bonds with His413 and Lys494 at this state. Because of the lower energy of the folded conformations, ATP shrinks back to its folded conformations, leading to the rupture of the hydrogen bond between ATP and Lys494. Consequently, Lys494 moves away from the pocket entrance, resulting in an open of the ATP lid of P4. It is the open state of P4 that can bind tightly to P1, where the His45 of P1 occupies a favorable position for its autophosphorylation from ATP. This indicates that ATP is not only a phosphoryl group donor but also an activator for CheA phosphorylation. Accordingly, a mechanism of the autophosphorylation of CheA is proposed as that the ATP conformational switch triggers the opening of the ATP lid of P4, leading to P1 binding tightly, and subsequently autophosphorylation from ATP to P1.     

Optimisation of sample pre-treatment method for the determination of triclosan in marine sediments by high-performance liquid chromatography and marine benthic quality assessment in the southern Baltic Sea

This study comprises optimisation of sample preparation and HPLC analytical procedure for the determination of a personal care product ingredient, triclosan (TCS), in marine sediments. The testing of several varying pre-treatment parameters confirmed that ultrasonic extraction is an effective method for the isolation of TCS from marine sediments, and that the choice of extraction solvent appeared to be of major importance. The selection of the mobile-phase composition and the absorption wavelength was made for the high-performance liquid chromatography analysis step. Based on the validated method, a preliminary assessment of the benthic ecosystem quality with regards to TCS contamination has been demonstrated in the southern Baltic Sea – a semi-enclosed sea, characterised by poor water exchange, thus particularly susceptible to anthropopression. TCS has been identified and quantified in situ in marine bottom sediments, sediment dwelling isopod – Saduria entomon L. and estimated in silico in pore waters based on the equilibrium partition theory in order to assess the potential exposure and uptake from the aqueous phase. TCS concentrations identified in the bottom sediments of the Gdansk Basin, as the natural habitat for studied S.entomon L., appear to be threatening to the benthic environment. Particularly when considering S. entomon L. as a major nutrition source for cod (Gadus morhua) undergoing the feminisation process, since the recent studies prove TCS to have a potential to induce critical alterations in the endocrine system of marine ichthyofauna.     

A semi-empirical method for calculation of true coincidence corrections for the case of a close-in detection in γ-ray spectrometry

In this paper, a semi-empirical method is proposed to determine true coincidence-summing (TCS) correction factors for high resolution γ-ray spectrometry. It needs the knowledge of both full energy peak (FEP) efficiency and total-to-peak (TTP) efficiency curves. The TTP efficiency curve is established from the measurements with a set of coincidence-free point sources. Whereas for a volume source, the coincidence-free FEP efficiency curve is obtained iteratively by using the peaks from almost the coincidence-free nuclides and those from the coincident nuclides in the mixed standard sources. Then the fitting parameters obtained for both TTP and FEP efficiency curves are combined in a freely-available TCS calculation program called TrueCoinc, which yields the TCS correction factors required for any nuclide. As an application, the TCS correction factors were determined for the particular peaks of ²³⁸U, ²²⁶Ra and ²³²Th in the reference materials, measured in the case of a close-in detection geometry using a well-type Ge detector. The present TCS correction method can be applied without difficulty to all Ge detectors for any coincident nuclide.     

Trichosanthin induced calcium-dependent generation of reactive oxygen species in human choriocarcinoma cells

The type-I ribosome-inactivating protein trichosanthin (TCS) has a broad spectrum of biological and pharmacological activities, including abortifacient, anti-tumor and anti-HIV. We found for the first time that TCS induced the generation of reactive oxygen species (ROS) in human choriocarcinoma cells (JAR cells) at the level of the single cell by using the fluorescent probe 2',7'-dichlorofluorescein diacetate with confocal laser scanning microscopy. TCS-induced ROS formation was shown to be dependent on the presence of extracellular Ca2+ and was further reduced when cytosolic Ca2+ was chelated by BAPTA-AM. The production of ROS increased rapidly after the application of TCS, which paralleled TCS-induced increase in intracellular calcium monitored using fluo 3-AM. Simultaneous observation of the nuclear morphological changes via two-photon laser scanning microscopy and production of ROS via confocal laser scanning microscopy revealed that ROS were involved in the apoptosis of JAR cells. The contribution of ROS was confirmed by experiments in which the antioxidant alpha-tocopherol prevented TCS-induced ROS formation and cell death. The finding that TCS induced calcium-dependent generation of ROS in JAR cells and that ROS were involved in the apoptosis of JAR cells might provide new insight into the anti-tumor and anti-HIV mechanism of TCS.