用荧光法研究维生素B6与白蛋白的相互作用

本文来分析用荧光碎灭法研究维生素Ｂ_６与人血清白蛋白和牛血清白蛋白间的相互作用，并对作用机理进行了分析．     

马兜铃酸A与牛血清白蛋白相互作用的研究

应用光谱法和电化学方法研究了水溶液中马兜铃酸A与牛血清白蛋白分子间的结合反应,测定了反应的形成常数KA及热力学函数ΔG,ΔH和ΔS,并确定了分子间作用力性质;还讨论了温度和微量金属离子对药物与牛血清白蛋白形成常数的影响;采用同步荧光技术考察了马兜铃酸A对牛血清白蛋白构象的影响;依据Forster非辐射能量转移机制,确定了给体—受体间的结合距离和能量转移效率.     

Hg~(2+)与NBS修饰前后两种血清蛋白结合的研究(英文)

外加毫摩尔浓度的重金属离子Hg2+,会引起BSA或HSA的色氨酸荧光下降,并且35℃时的变化比25℃时更明显,HSA的变化比BSA的大.SternVol mer作图分析结果表明荧光淬灭主要是由动态淬灭引起的.用NBS对BSA和HSA的色氨酸残基进行特异性修饰后,Hg2+只引起NBS修饰的HSA的酪氨酸荧光轻微降低,但对NBS修饰的BSA的酪氨酸荧光几乎没有影响.远紫外和近紫外CD谱分析表明:Hg2+的结合要引起BSA,HSA以及NBS修饰的HSA二级结构的α螺旋减少,三级结构发生一定的改变.这些结果表明B     

牦牛血清转铁蛋白对人红细胞膜氧化损伤的保护作用

牦牛血清中分离纯化的转铁蛋白对人红细胞膜的抗氧化具有保护作用 .方法 :采用黄嘌呤 /黄嘌呤氧化酶、Fe2 + /H2 O2 两个体系产生的O·2 -、OH·对红细胞膜造成氧化损伤 ,通过丙二醛含量、超氧化物歧化酶和谷胱甘肽过氧化酶活性的测定研究牦牛转铁蛋白在抗氧化方面的作用与机制 .结果 :O·2 -和OH·可以造成红细胞膜丙二醛量的显著增加 ,超氧化物歧化酶和谷胱甘肽过氧化酶活性的下降 .当适量不同浓度牦牛血清转铁蛋白在黄嘌呤 /黄嘌呤氧化酶、Fe2 + /H2 O2 两个体系前处理可以抑制丙二醛的产生 ,保护红细胞膜免受丙二醛的氧化损伤并且使超氧化物歧化酶和谷胱甘肽过氧化酶活性升高 .提示 ,牦牛血清转铁蛋白对O·2 -和OH·自由基引起的氧化损伤有保护作用     

银纳米粒子与血清白蛋白相互作用的滞后效应

用圆二色光谱,紫外-可见光谱研究银纳米粒子与血清白蛋白的相互作用及其相关效应.拟合计算圆二色光谱数据发现,与银纳米粒子作用后的血清白蛋白中α-螺旋含量减少,而α-折叠、转角和无规卷曲等结构含量增加,其构象在作用后变得更为松散和伸展.通过时间扫描吸收光谱跟踪研究发现,在血清白蛋白与银纳米粒子相互作用的过程中,银纳米粒子的包覆与聚集、血清白蛋白的构象变化具有双重滞后效应.     

Effect of bovine serum albumin on the surface properties of ionic liquid-type Gemini surfactant

The effect of bovine serum albumin on the surface properties of IL-type gemini surfactant ([C₁₀-4-C₁₀im]Br₂), have been investigated by surface tension method. The critical micelle concentration (CMC) as a function of BSA concentrations at various temperatures was investigated. The CMC of [C₁₀-4-C₁₀im]Br₂ increases with increasing the concentration of BSA as well as the temperature of the system. The interfacial parameters viz; maximum surface excess concentration (Γ_max), the minimum area per molecule (A_min), and surface pressure at CMC (Π_cmc) were calculated. In addition, thermodynamic parameters of adsorption and micellization were evaluated by using surface tension data. The results indicated that the binding of [C₁₀-4-C₁₀im]Br₂ to BSA is spontaneous and exothermic in nature. The process is entropy driven and hydrophobic interactions are the major driving forces.     

Switchable peptide-equipped protein/cucurbit[7]uril supramolecular assembly for targeted drug delivery

ABSTRACT

Herein, we develop a switchable peptide-equipped protein/cucurbit[7]uril (CB[7]) supramolecular assembly as novel targeted drug vector. Specifically, bovine serum albumin (BSA) is used to interact with CB[7], serving as the core of drug vector. Then, a peptide shield layer is formed on the surface of BSA/CB[7], yielding peptide-equipped supramolecular assembly (Pep@BSA@CB[7]). The equipped peptide shield layer is composed of switchable peptide probes consisting of a polycationic cell-penetrating peptide (CPP) motif, a polyanionic motif and a linking motif, and therefore provides a variety of desirable properties. First, the CPP motif displays excellent cell penetration ability and can facilitate internalisation of the drug vector. Secondly, the polyanionic motif performs intramolecular electrostatic interaction with CPP motif and thereby can reduce non-targeted delivery towards normal cells. Thirdly, the linking motif can be specifically cleaved by matrix metalloproteinases 2 that is up-regulated in tumour microenvironment, thus enabling precise cancer-targeting. As a consequent, Pep@BSA@CB[7] can serve as a promising drug vector that exhibits superior targeting ability and high uptake efficiency towards cancer cells, which may be of great potential in cancer-targeted treatment.     

Cyclic sample pooling using two‐dimensional liquid chromatography system enhances coverage in shotgun proteomics

We report a cyclic sample pooling technique devised in two‐dimensional liquid chromatography–electrospray ionization mass spectrometry (LC‐ESI‐MS) shotgun proteomics that renders deeper proteome coverage; we combined low pH reversed‐phase (RP) LC in trifluoroacetic acid in the first dimension, followed by cyclic sample pooling of the eluate and low‐pH RP‐LC in formic acid in the second dimension. The new protocol has a significantly higher resolving power suitable for LC‐ESI‐MS/MS shotgun proteomics. Copyright © 2013 John Wiley & Sons, Ltd.     

Development and optimization of ultra-high performance supercritical fluid chromatography mass spectrometry method for high-throughput determination of tocopherols and tocotrienols in human serum

The goal of this study was to develop an effective supercritical fluid chromatography method using single quadrupole MS for analysis of all isomeric forms of vitamin E. Finally, two fast and effective methods, the high resolution one and the high speed one, for the determination of 8 vitamin E isomers in human serum were developed.     

Colorimetric and Spectrofluorimetric Methods for the Determination of Melatonin in Tablets and Serum

Abstract

Two sensitive and accurate colorimetric and spectrofluonmetric methods, are presented for the determination of melatonin in tablets and serum. The first method utilizes the reactions of p-dimethylaminobenzaldehyde in hydrochloric acid (van Urk reagent)-ferric chloride in sulphuric acid (Salkowski reagent) mixture. The blue color of the resulting reaction product is measured at 630 nm. The second method is based on the reaction of melatonin with o-phthalaldehyde in acid medium which yields highly fluorescent condensation product that is measured at 465 nm as emission wavelength, using excitation wavelength at 355 nm. No interference was observed from tableting additives, and the applicability of the methods was examined by analysing tablets containing melatonin (single and combined with pyridoxine). Mean percentage recoveries from tablets were found to be 99.9+0.31 for single and 100.5+0.15 for combined tablets using colorimetric method, while by applying spectrofluorimetric method the recoveries were found to be 100.610.41 for single and 100.2+0.39 for combined tablets. Furthermore, the proposed methods were extended to the in-vitro determination of melatonin in serum. The detection limits are 0.27 ug ml^?1 for colorimetric method and 0.00035 ug ml^?1 for spectrofluorimetric method.