Study of the Interaction of Hemoglobin with Ligands from Resonance Raman Spectra

By the method of resonance Raman spectroscopy the equilibrium between liganded (HbL) and deliganded (Hb) forms of human hemoglobin is studied in destabilization of the solvate protein shell by introducing a cosolvent in the solution. It is shown that the introduction of ethanol leads to a nearly fivefold decrease in the ligand affinity of hemoglobin detected from the increase in the intensity of the 1355 cm^–1 ₄ line. The conformation of the heme and of the hemic pocket remains constant on addition of ethanol to the solution. Owing to the short time spent on measurement, this method can be used for a rapid evaluation of the influence of various preparations on the ligand affinity of hemoglobin.     

A Hydrogen Peroxide Biosensor Based on Room Temperature Ionic Liquid Functionalized Graphene Modified Carbon Ceramic Electrode

A room temperature ionic liquid (IL) 1‐butyl‐3‐methylimidazolium hexafluorophosphate functionalized graphene (GE) was prepared and a hydrogen peroxide (H₂O₂) biosensor was fabricated by immobilizing hemoglobin (Hb) into the IL‐GE composite film. UV‐visible and Fourier transform infrared spectra of the composite film indicated that Hb retained its native structure in the film. Electrochemical investigation of the biosensor showed a pair of well‐defined, quasi‐reversible redox peaks with E_pa=?0.209 V and E_pc= ?0.302 V (vs. SCE) in pH 7.0 phosphate buffer solution at the scan rate of 100 mV/s. To the reduction of H₂O₂, the biosensor had a good linear range from 8.0×10^?7 to 1.8×10^?4 mol/L with a detection limit of 3.0×10^?7 mol/L. The apparent Michaelis‐Menten constant K^app_M was estimated to be 3.4×10^?5 mol/L.     

Structure Characteristic and Catalase Activity of Vitreoscilla Hemoglobin Bound with Membrane

Lipid-bound [VHb(406)] and lipid-free [VHb(402)] wide type Vitreoscilla hemoglobin were separated from E. coli BL21(DE3). The RZ is 3.30 for [VHb(406)] and 3.15 for [VHb(402)], respectively. VHb(406) shows a characteristic absorption band at 626 nm, while VHb(402) shows one at 644 nm, and it has more α-helix than VHb(402). Data of Raman spectra experiment shows under the excitation wavelength 488 nm, υ4 vibration of both VHb(402) and VHb(406) had a pure and strong signal at 1375 cm-1, which proves that iron...     

Preintercalation of Layered γ—Zirconium Phosphate for Preparation of Immobilized Hemoglobin

Layered γ-Zirconium phosphate (γ-ZrP) preintercalated with butylamine,tetra(n-butylammonium)hydroxide,dimethylamine respectively,or only ultrasonificated,for preparation of immobilized hemoglobin were in vestigated in this report.     

浙江萧山、舟山和福建集美臭鼩血红蛋白、血清蛋白电泳比较研究

本文用血红蛋白的薄层等电聚焦技术对浙江萧山、舟山和福建集美的三地臭鼩进行比较,结果显示:大陆(萧山与集美)和海岛(舟山)之间的带谱存在差异.大陆有6条区带,海岛的带谱变化较大,存在四种不同类型.用血清蛋白的SDS-聚丙烯酰胺凝胶电泳技术和血清蛋白的薄层等电聚焦技术对上述三地臭鼩的分析表明:海岛个体血清蛋白的成份和在血液中相对含量较大陆为少,而大陆萧山和集美的血清蛋白在血液中的相对含量和成份亦略有不同.     

不同pH对血红蛋白与过氧化氢复合物中间体的影响

以血红蛋白模拟过氧化物酶，在不同pH溶液中，测定并比较了催化反应中间体－Ⅱ的ks,t1/2和催化活性等。结果表明，中间体－Ⅱ的ks,t1/2和催化活性均与缓冲液的pH密切相关，其稳定性在pH5.0时最小，而此时的催化反应速度最高。     

嗜中性粒细胞中过氧化氢及其释放过程的电化学检测

活性氧（ROS）是细胞代谢的重要产物之一,其浓度与多种生理、病理现象密切相关;H2O2是ROS的重要代表,对细胞内H2O2及其释放过程的测定有助于理解相关疾病的形成机制,因而具有重要的意义.本文报道一种检测细胞内H2O2浓度、监测其生成和释放过程的电化学方法,该方法利用固定在天然矿物凹土（attapulgite,全称凹凸棒石黏土）表面的血红蛋白（Hb）对H2O2还原的电催化作用实现H2O2的定量测定.将Hb固定到凹土表面制备了Hb-凹土（Hb-Atta）纳米复合材料,用透射电子显微镜（TEM）、原子力显微镜（AFM）、红外光谱（FTIR）、紫外-可见光谱（UV-vis）等对其进行了表征;将该复合物修饰到玻碳（GC）电极表面制成了Hb-Atta/GC电极,用伏安法考察了Hb在凹土表面的直接电子转移性能,结果表明,Hb能进行有效的直接电子转移反应,其伏安曲线上呈现出一对良好的氧化还原峰,式量电位（E0′）约为-350mV（vs.SCE,50mV/s）;进一步的结果表明,Hb-Atta对H2O2的电化学还原具有良好的催化性能,并能用于H2O2的定量测定,具有响应速率快、线性范围宽、检测限低等优点.将Hb-Atta/GC电极应用于嗜中性粒细胞内H2O2的测定,结果表明,该电极不仅能定量地检测细胞内的H2O2含量,还能监测细胞内H2O2生成和释放的过程,而且细胞内其他ROS和电活性物质如OCl-、NO.、ONOO-和抗坏血酸等不产生干扰.本文的结果为测量细胞内H2O2浓度、研究其生成及释放过程动力学以及进一步阐明由H2O2导致的相关疾病机制等提供了一个新的平台,可望在生物电化学、细胞生物学和病理生理学等领域得到应用.     

Interaction of Moxifloxacin with Bovine Hemoglobin and Effect of the Coexistent Drugs on the Reaction

The interaction between moxifloxacin (MXFX) and Bovine Hemoglobin (BHb) was investigated at different temperatures by fluorescence spectroscopy. Results showed that the quenching mechanism of MXFX on BHb was a static quenching process with Förester spectroscopy energy transfer. The primary binding for MXFX was located at β-37 Tryptophan residue in hydrophobic cavity of BHb. Besides, weak negative cooperativity was found in drug's binding with BHb. Synchronous spectra revealed that the microenvironment and the conformation of serum albumin were changed during the reaction. Most antibiotics had no effect on the system of BHb-MXFX, except quinolone antibiotics. The system had good stability.     

空间和地面条件下斑头雁血红蛋白晶体生长母液的共振拉曼光谱研究

对在宇宙飞船和地面进行斑头雁血红蛋白晶体的生长后保留的母液进行了共振拉曼光谱的比较研究。通过分析两种样品在1240cm-1和1360cm-1附近的共振拉曼信号的细微差别,我们认为空间样品中Fe和N的配位强度平均要比地面样品大,并推测这是由空间微重力环境所致。