Electrochemically Induced Iron Release of Adsorbed Horse Spleen Ferritin: Quantitation of Iron Using Long Optical Path Length Thin‐Layer Spectroelectrochemistry

In this work, long optical path length thin‐layer electrochemical cell was constructed using indium‐tin oxide on glass as the electrode material. Iron release from ferritin adsorbed on the electrode was induced by applying a negative potential sweep in the presence of 1,10‐phenanthroline. The usefulness of spectroelectrochemistry as a means of determining the quantity of iron released from an adsorbed layer of ferritin is demonstrated.     

A method for C(alpha) direct-detection in protonless NMR

Attempts are made to efficiently decouple (13)C nuclei without significant loss of coherence during the application of the decoupling package. Such attempts are based on the S(3)E spin-state selection method. A newly developed double S(3)E (DS(3)E) is particularly efficient for C(alpha) detection for proteins as large as 480 kDa.     

Electron holography of biological samples

In this paper, we summarise the development of off-axis electron holography on biological samples starting in 1986 with the first results on ferritin from the group of Tonomura. In the middle of the 1990s strong interest was evoked, but then stagnation took place because the results obtained at that stage did not reach the contrast and the resolution achieved by conventional electron microscopy.

To date, there exist only a few (12) publications on electron holography of biological objects, thus this topic is quite small and concise. The reason for this could be that holography is mostly established in materials science by physicists. Therefore, applications for off-axis holography were powerfully pushed forward in the area of imaging, e.g. electric or magnetic micro- and nanofields. Unstained biological systems investigated by means of off-axis electron holography up to now are ferritin, tobacco mosaic virus, a bacterial flagellum, T5 bacteriophage virus, hexagonal packed intermediate layer of bacteria and the Semliki Forest virus. New results of the authors on collagen fibres and surface layer of bacteria, the so-called S-layer 2D crystal lattice are presented in this review. For the sake of completeness, we will shortly discuss in-line holography of biological samples and off-axis holography of materials related to biological systems, such as biomaterial composites or magnetotactic bacteria.     

鲨鱼肝铁蛋白亚基解离与组装机理的研究

小批量制备质谱纯鲨鱼肝铁蛋白(Liver Ferritin of Sphyrna zygaena,SZLF)。在弱酸介质(pH1.0)中,天然电泳结果显示,SZLF蛋白质亚基20 min后开始解离。选用透射电子显微镜跟踪SZLF亚基解离与重组装全过程和蛋白壳与铁核尺寸变化,发现SZLF在亚基酸解离过程中,随着pH值的降低,铁核和蛋白壳的尺寸呈现相同的变化趋势,这种变化趋势可能与铁核内层铁的释放和蛋白壳的解离与去折叠有关。SZLF蛋白壳的重组装过程则是一个快速过程,并且是由松散熔球态向紧密态转变的过程。SZLF由单类型亚基组成,而马脾铁蛋白(Horse Spleen Ferritin,HSF)由H和L两种亚基类型组成。在基质pH3.0条件和激光辅助下,混合HSF和SZLF仍然可释放各自的亚基且形成准亚基离子,供基质辅助激光解析电离飞行时间质谱分析,说明此时SZLF的亚基间相互作用强度减弱但并没有去折叠。TEM技术在铁蛋白解离和重组装过程中的应用,为进一步研究铁蛋白纳米包装的过程和机理提供新颖的、可行的和更加直观的研究手段。     

黄豆铁蛋白提取新方法及其与豌豆铁蛋白活性比较

铁蛋白是广泛存在于动物、微生物及植物体中的一种铁贮藏蛋白, 具有去除二价铁的毒性以及调节机体细胞铁代谢平衡的作用. 本文以黄豆种子为原料, 开发出黄豆铁蛋白提取新方法, 即将黄豆粗提液在55 ℃下加热15 min, 再将冷却后的上清液分别用500 mmol/L MgCl2和700 mmol/L柠檬酸三钠进行盐析. 离心得到的铁蛋白粗提液经透析后, 用DEAE-cellulose弱阴离子交换层析和Sephacryl S-300凝胶过滤层析进一步分离, 得到电泳纯的铁蛋白. Native-PAGE电泳测得分子量约为560000, SDS-PAGE电泳结果表明, 黄豆铁蛋白含有两种亚基, 分子量分别为28000和26500. 活性研究显示, 黄豆铁蛋白与豌豆铁蛋白铁氧化沉淀和还原释放反应的活性明显不同.     

用圆二色性和荧光光谱技术研究纳米吡啰红G核-铁蛋白的构建机理

小批量制备了电泳纯鲨鱼肝铁蛋白(Liver ferritin ofSphyrna zygaena,SZLF),对SZLF的结构与功能进行了研究.在pH=2.0~10.0条件下,选用圆二色性(Circular dichroism,CD)光谱技术研究了SZLF和脱铁核SZLF(apoSZLF)二级结构转换的基本特征和变化趋势,揭示了SZLF蛋白壳的亚基稳定性、相互作用强度和去折叠现象.利用酸碱中和方法,解离SZLF蛋白壳亚基,并再次构建成完整SZLF.用紫外-可见分光光度法和荧光光度法研究了构建纳米吡啰红G核-铁蛋白的途径.定量分析结果表明,每分子apoSZLF可捕获12分子吡啰红G于蛋白壳内,构建纳米吡啰红G核-铁蛋白.分析了apoSZLF直接捕获和释放吡啰红G的途径与速率,为后续构建高存量纳米顺铂核-铁蛋白药物载体提供了可行性技术.     

Excitation of plant growth in dormant temperature by steady magnetic field

Experimental results of applying a steady magnetic field (20 and 30 mT) on agricultural plants reveal that their growth is more than that of control plants. Considering that these plants have ferritin cells, and each ferritin cell has 4500 Fe atoms, it is obvious that they have an outstanding role in the plants’ growth. As the last spin magnetic moment (SMM) of the Fe atom posed to an external magnetic field (EMF), the composition of SMM and EMF create an oscillator in the system. Then we have a moment of force on ferritin cells. This oscillator exerts its energy, then damps and finally locates in the field direction. The relaxed energy increased the internal temperature (i.e., the effective temperature of the magnetic spin system of plant) so that it is situated in a proper temperature for growing. This phenomenon (temperature increasing) occurs in the initial minutes of applying the magnetic field. So it depends on the number of times of locating the plant in magnetic field in a day (n). If this number (n) passes the critical value, the plant reaches a burning temperature and growth is perturbed. In this paper, the plant growth rate and critical temperature in a steady magnetic field were investigated and formulated theoretically. An innovative result in this research is as follows: if a plant's environment was in the dormant temperature, we could increase the internal temperature of the plant by applying a magnetic field n times in a day (for growth).     

Redox reaction characteristics of ferritin-immobilized onto poly(l-lysine)-modified indium oxide electrodes

Ferritin-immobilized poly(l-lysine)-modified electrodes showed well-defined redox waves representing ferritin. Cathodic and anodic peak currents obtained from cyclic voltammograms were proportional to potential sweep rates. From charge flow values during oxidation or reduction reactions calculated by peak areas in cyclic voltammograms, and the surface coverage of ferritin, reacted iron atoms per ferritin molecule were calculated. Obtained numbers of reacted iron atoms were significantly smaller than expected values from iron atoms at ferrihydrite core surfaces of ferritin, which would be caused by the rate-determining ion flow through ion channels of ferritin to compensate for charges in the ferritin cavity. Anodic and cathodic peak potentials in cyclic voltammograms were significantly dependent on cationic species in the solution, though voltammetric shapes and peak currents were independent of cations. From the obtained results that structural changes in ferritin were not detected by fluorescent spectra, it is thought that the cationic dependence on ferritin redox peak potentials is caused by ferritin cores.     

Protective role of exogenous nitric oxide against oxidative-stress induced by salt stress in barley (Hordeum vulgare)

To probe into the potential of relieving the oxidative damage of salt stress, we investigated the protective role of nitric oxide on barley under salt stress. Salt stress resulted in increased ion leakage, lipid peroxidation and protein oxidation in barley leaves. Simultaneous treatments of barley leaves with 50 microM sodium nitroprusside, a nitric oxide donor, alleviated the damage of salt stress, reflected by decreased ion leakage, and malendialdehyde (MDA), carbonyl, and hydrogen peroxide content in barley leaves. The presence of the nitric oxide donor increased the activities of superoxide dismutases (SOD), ascorbate peroxidases (APX), and catalases (CAT). Meantime, sodium nitroprusside addition increased accumulation of ferritin at the protein level, indicating that nitric oxide directly regulated ferritin accumulation. These results suggested that nitric oxide can effectively protect seedlings from salt stress damage by enhancing activities of antioxidant enzymes to quench the excessive reactive oxygen species caused by salt stress and inducing the increase of ferritin accumulation to chelate larger number of ferrous ion. Information from this study can be used to improve soil management practices for sustainable use of salt-affected soils in the future.     

原子吸收光谱法检测血清铁、红细胞内铁及血清铁蛋白、 MCH、MCHC含量与老年冠心病、高血压病关系的研究

应用原子吸收光谱法检测了老年高血压、冠心病患者血清铁、红细胞内铁及血清铁蛋白、ＭＣＨ、ＭＣＨＣ。发现老年冠心病患者血清铁、红细胞内铁、血清铁蛋白含量明显高于对照组（Ｐ＜０．０１）。老年高血压患者血清铁蛋白、红细胞内铁、ＭＣＨ、ＭＣＨＣ也明显高于对照组（Ｐ＜０．０１）,血清铁与对照组比较有显著性差异（Ｐ＜０．０５）。结果表明：老年高血压,冠心病患者存在铁代谢异常,其游离铁与储存铁的增加可能是其发病的危险因素之一。