Fluorescence of an eosine probe in solutions of human serum albumin with organic and inorganic ligands

The fluorescence of an eosine molecular probe in solutions of human serum albumin (HSA) in the presence of an inorganic (CsCl) and organic (sodium dodecyl sulfate, SDS) ligand was studied. The interaction of HSA molecules with these ligands was analyzed. It was demonstrated that the presence of CsCl, a low-molecular-weight salt, in the solutions influences the efficiency of the complexation of eosine with HSA. Data on the dynamics of the conformation rearrangement of HSA during denaturation under the action of SDS were obtained.     

离子对人血清白蛋白影响的1H NMR研究

人血清白蛋白（HSA）上有很多的结合位点,对药物及内源性物质的运输有很重要的作用.由于HSA分子量大且结构复杂,核磁共振（NMR）谱图信号重叠严重,因此使用常规的NMR技术很难获得天然丰度HSA上结合位点的信息.我们通过新的谱编辑技术,将横向弛豫加权（T₂W）与RD-WaterLOGSY技术相结合（即T₂W-RD-WaterLOGSY）,观察到了天然丰度HSA表面的一些明显的特征信号,且这些信号的分辨率较高;通过pH滴定和2D ¹H-¹H TOCSY实验,我们对观察到的信号做了初步指认,发现部分信号来自于HSA表面的组氨酸;然后,我们通过这些特征信号研究了金属离子与HSA之间的相互作用.结果表明该技术可用于简化天然丰度HSA的谱线,在不对HSA进行突变的情况下,能反映了Zn²⁺与HSA的强结合作用以及结合位点等信息.     

Structural Changes in Human Serum Albumin According to the Data on the Phosphorescence Kinetics of a Luminescent Probe — Eosin

The influence of the human serum albumin (HSA) denaturation by a surface-active substance (sodium dodecyl sulfate (SDS)) on the phosphorescence of a luminescent probe (eosin) has been investigated. The dependences of the eosin-phosphorescence intensity on the SDS concentration were determined at different pH levels of an HSA solution. It has been shown that at SDS concentrations lower than the critical concentration necessary for micelle formation, the hydrophobic interactions of eosin with the protein influence the deactivation of the eosin triplet states. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 72, No. 5, pp. 660–663, September–October, 2005.     

The fluorescence characteristics and the molecular association of the Rose Bengal nanomarker in human serum albumin solutions

The decrease in the degree of molecular association of the Rose Bengal nanomarker in solutions with the addition of human serum albumin (HSA) has been revealed. It has been observed that in solutions with the addition of HSA the fluorescence quenching and the shifting of the fluorescence spectrum peaks of Rose Bengal to the red take place. It has been shown that the dependence of the effective binding constant of binding Rose Bengal to HSA steadily decreases with an increase in the pH value. It has been established that the values of the molecular association degree of Rose Bengal and the values of the effective constant of its binding to HSA depend on the magnitude of the electronegativity of the atoms in its structural formula, as well as on the pK values of its ionizable groups.     

铽标记人血清白蛋白的荧光光谱及应用

本文研究了由对-氨基水杨酸衍生的铽资合物及其标记人血清白蛋白的荧光光谱;测量了它们的激发态寿命.首次提出铽资合物标记蛋白质中蛋白质对Tb3+离子发光无影响.建立了液相中竞争性的人血清白蛋白的荧光免疫分析法.     

Structure characterization and stability of mixed lipid/protein monolayer at the air/water interface

A mixed protein/lipid monolayer has been constructed by the protein adsorption from subphase into the spread phospholipid monolayer. A precisely controlled pump was used to exchange the protein solution with different pH values after the protein was ensured to reach the less condensed surface.

The domains formed in the coexistence region of D-dipalmitoylphosphatidylcholine (D-DPPC) have been recorded by Brewster angle microscopy (BAM) combined with the film balance before and after the penetration of the protein, human serum albumin (HSA). The subphase was exchanged by gradually increasing or decreasing pH value of the solution. Three isotherms of the mixed D-DPPC/HSA monolayer with the subphase of pH=4.2, pH=7.0 and pH=9.1, respectively, were obtained. It indicated that the area per lipid molecule with protein increased as the subphase pH value was lowed. Simultaneously, morphological dynamic changes caused by the gradual changing on subphase pH were observed. These variations can be ascribed to the conformation change of protein under the fluctuation of pH value. The hydrophobic and electrostatic interactions between the phospholipid and HSA were as considered for the interpretation of domain change based on the current experimental results     

基于RRS的人血清白蛋白快速检测系统设计

在临床中人血清白蛋白浓度的快速检测对多种重大疾病的诊断具有重要意义,而现有的医疗检测设备普遍存在体积大、测试周期长、操作复杂等缺点,设计研究一种快速的、高精度的便携式人血清白蛋白浓度检测系统具有非常广阔的应用前景。设计了基于共振瑞利散射光谱分析的人血清白蛋白浓度检测系统。由于人血清白蛋白与四氨基酞菁铜反应产生的溶液在475.0 nm处具有共振瑞利散射增强的效果,从而构建了共振瑞利散射强度与人血清白蛋白浓度的函数。实验采用紫外半导体激光器与475.0 nm窄带滤光片联用,配合高增益光电探测器采集散射光信号。采用不同浓度的四氨基酞菁铜溶液分别对不同浓度的人血清白蛋白进行检测,实验结果显示,响应电压会随人血清白蛋白浓度的提高而明显增大,而对四氨基酞菁铜的浓度变化并不敏感;散射光产生的响应电压与人血清白蛋白浓度在0.1～1.0 μg·mL-1范围内基本满足线性变化,满足快速检测的设计要求。     

Comparative analysis of cesium and potassium uptake in onion <Emphasis Type="Italic">Allium cepa</Emphasis> L.

Cesium uptake in onion (from 0.3 mM CsCl solution traced with ¹³⁷CsCl) has been examined. The highest uptake occurred at pH 4–5 and it decreased with increasing pH. The intensity of Cs translocation depended on acidity of the solution. For acidic solutions, translocation of cesium into bulbs and leaves was greater than in case of alkaline solutions, where most of the cesium remained in the roots. Addition of potassium into the solutions (millimolar K concentrations) inhibits Cs uptake. The potassium pH-influx/efflux characteristic does not coincide with the Cs uptake.     

A study of the adsorption of the amphiphilic penicillins cloxacillin and dicloxacillin onto human serum albumin using surface tension isotherms

The interaction of human serum albumin (HSA) with two structurally similar anionic amphiphilic penicillins, cloxacillin and dicloxacillin, at 25 °C has been examined by surface tension measurements under conditions at which the HSA molecule was positively (pH 4.5) or negatively charged (pH 7.4). Measurements were at fixed HSA concentrations (0.0125 and 0.125% w/v) and at drug concentrations over a range including, where possible, the critical micelle concentration (cmc). Interaction between anionic drugs and positively charged HSA at pH 7.4 resulted in an increase of the cmc of each drug as a consequence of its removal from solution by adsorption. Limited data for cloxacillin at pH 4.5 indicated an apparent decrease of the cmc in the presence of HSA suggesting a facilitation of the aggregation by association with the protein. Changes in the surface tension-log (drug concentration) plots in the presence of HSA have been discussed in terms of the adsorption of drug at the air-solution and protein-solution interfaces. Standard free energy changes associated with the micellization of both drugs and their adsorption at the air-solution interface have been calculated and compared.     

荧光法研究PTB与白蛋白的结合

利用药物对蛋白的荧光猝灭作用,用荧光法研究了N-苯酰甲噻唑溴(PTB)与牛血清白蛋白(BSA)及人血清白蛋白(HSA)的相互作用。测定发现BSA溶液的最大激发波长为280 nm,HSA溶液的最大激发波长为290 nm。分别向溶液中加入PTB后,原有的最大发射波长处的强度明显减弱。说明PTB对BSA和HAS有荧光猝灭作用。PTB与BSA,HSA有中等强度的结合。测得15 ℃时PTB与BSA,HSA的结合常数分别为3.66×103和3.83×103,结合位点数n分别为1.02和1.16;37 ℃时PTB与BSA,HSA的结合常数分别为3.58×103和3.35×103,结合位点数分别为0.95和0.87。根据热力学常数确定了PTB与BSA,HSA之间的主要作用力类型均为静电作用力。通过Fster偶级-偶级非辐射能量转移原理,得到BSA,HSA与PTB结合的位置距色氨酸残基的距离分别为7.5和7.9 nm。根据白蛋白的结构,可以推测BSA,HSA与PTB结合的位点在ⅡA亚结构域,靠近Try214的区域。     

HSA-磷钼杂多酸缔合纳米微粒体系荧光猝灭机理研究

在pH 7.40的Tris-HCl缓冲溶液中, 磷钼杂多酸(PMA)呈浅黄色,在可见光区没有明显的吸收峰;人血清白蛋白(HSA) 呈无色,在可见光区也没有明显的吸收峰,但在350 nm处有一荧光峰。当有PMA存在时,HSA与PMA形成缔合纳米微粒, HSA-PMA缔合纳米微粒的粒径约为80 nm。经研究发现HSA对PMA有增色和减色效应,PMA对HSA有荧光猝灭作用;PMA对色氨酸(Trp)和酪氨酸(Tyr)也有一定的荧光猝灭作用,但这两种荧光猝灭机理不同,且没有形成缔合纳米微粒。PMA对色氨酸(Trp)和酪氨酸的荧光猝灭作用主要是由于PMA 在发射波长范围内存在一定的分子吸收,即通常所报道过的能量转移所至。研究结果表明,HSA-PMA缔合纳米微粒和界面的形成是导致该体系的荧光猝灭、共振散射增强及增色和减色效应的根本原因。     

Mn（Ⅱ）,Co（Ⅱ）与HSA相互作用的荧光光谱研究

用荧光光谱法研究了生理ｐＨ和等离子点（ｐＨ＝５．３０）时Ｍｎ（Ⅱ）、Ｃｏ（Ⅱ）与ＨＳＡ的相互作用。根据Ｆｏｒｓｔｅ非辐射能量转移理论,得到了不同ｐＨ时Ｍｎ（Ⅱ）、Ｃｏ（Ⅱ）在ＨＳＡ中的第一强结合位置与Ｔｒｐ－２１４残基间的距离。这一结果远大于文献报道值,根据Ｍｎ（Ⅱ）、Ｃｏ（Ⅱ）在ＨＳＡ中的结合部位及ＨＳＡ的畴结构对这一显著差异进行了讨论。     

氯化铯甲醇溶液对有机聚合物太阳能电池的影响

研究了氯化铯的甲醇溶液作为阴极修饰层,来提高传统有机聚合物太阳能电池器件性能。通过电容-电压(C-V)测试分析了铝电极和PTB7/PC_(70)BM之间的界面电荷积累情况,同时测试了紫外光电子能谱(UPS),对铝的功函数改变作了研究。结果表明,采用氯化铯的甲醇溶液作阴极修饰层的器件,其短路电流(J_(sc))、开路电压(V_(oc))、填充因子(FF)都有所提高,光电转化效率达到6.36%,与仅用甲醇处理过的器件相比,光电转化效率提高了11%;与未经甲醇处理的器件相比,光电转化效率提高了42.6%。这种一步溶液处理法能够减少电荷积累,同时降低铝电极的功函数,利于电子收集,进而提高器件性能。     

Peculiarities of the Perturbation of Water Structure by Ions with Various Hydration in Concentrated Solutions of CaCl2, CsCl,KBr, and KI

The degree of water structuring in solutions of four salts (CsCl, KBr, and KI with a concentration of 1M and CaCl₂ with a concentration of 0.5M) has been analyzed using THz time-domain spectroscopy. It is shown that the degree of water structuring in a solution of salt, prepared based on a highly diluted aqueous solution of the same salt, differs in some cases from the degree of structuring of similar solutions prepared on the basis of highly diluted aqueous solutions of other salts and similarly prepared water. The degree of water structuring increases in salt solutions containing ions with pronounced positive hydration, decreases in salt solutions containing ions with pronounced negative hydration, and does not differs from control in salt solutions containing no ions with pronounced positive or negative hydration.

     

Human‐Serum‐Albumin‐Coated Prussian Blue Nanoparticles as pH‐/Thermotriggered Drug‐Delivery Vehicles for Cancer Thermochemotherapy

Constructing novel multimodal antitumor therapeutic nanoagents has attracted tremendous recent attention. In this work, a new drug‐delivery vehicle based on human‐serum‐albumin (HSA)‐coated Prussian blue nanoparticles (PB NPs) is synthesized. It is demonstrated that doxorubicin (DOX)/HSA is successfully loaded after in situ polymerization of dopamine onto PB NPs, and the PB@PDA/DOX/HSA NPs are highly compatible and stable in various physiological solutions. The NPs possess strong near‐infrared (NIR) absorbance, and excellent capability and stability of photothermal conversion for highly efficient photothermal therapy applications. Furthermore, a bimodal on‐demand drug release sensitively triggered by pH or NIR irradiation has been realized, resulting in a significant chemotherapeutic effect due to the preferential uptake and internalization of the NPs by cancer cells. Importantly, the thermochemotherapy efficacy of the NPs has been examined by a cell viability assay, revealing a remarkably superior synergistic anticancer effect over either monotherapy. Such multifunctional drug‐delivery systems composed of approved materials may have promising biomedical applications for antitumor therapy.     

Study of the Interaction of Fangchinoline with Human Serum Albumin by Fluorescence and UV Spectroscopic Method

The interaction of fangchinoline with human serum albumin (HSA) was studied by use of fluorescence quenching spectra, synchronous fluorescence spectra, and ultraviolet spectra. It was shown that fangchinoline has a strong ability to quench the fluorescence of HSA. The Stern‐Volmer curves based on the quenching of the fluorescence of HSA by fangchinoline indicated that the quenching mechanism of fangchinoline on HSA was static quenching and non‐radiation energy transfer. Based on the Förster theory of non‐radiation energy transfer, the binding distances (r) and the binding constants (K _A) between fangchinoline and HSA were found. The thermodynamic parameters obtained revealed that the interaction between fangchinoline and HSA was mainly driven by hydrophobic force. The conformational changes of HSA were investigated by use of synchronous fluorescence. The result indicates that an ionic electrostatic interaction between fangchinoline and HSA could not be excluded.     

利复星与人血清白蛋白作用的荧光光谱研究

用荧光光谱研究了药物利复星(Levofloxacin)与人血清白蛋白(HSA)的作用和影响,测量发现人血清白蛋白的最大激发峰位于286.70 nm处。在向该溶液滴加Levofloxacin时,原有的343.70 nm发射峰强度明显减弱, 且向长波长稍有移位,并出现了位于503.96 nm的新荧光发射峰(利复星的发射峰), 利复星对HSA荧光有猝灭作用。利复星Levofloxacin的503.96 nm荧光的激发峰则位于300.16和336.16 nm。当向该溶液滴加利复星时,300.16和336.16 nm的激发峰仅向长波长方向稍有移动。利复星对HSA的离解常数K_d=3.65×10-5(mol·L-1)。利复星的结合常数为K_S=2.742×104(L·mol-1)。利复星-HSA体系的猝灭过程不是因为分子扩散和碰撞所引起的动态猝灭,而是分子之间结合形成了化合物所引起的静态猝灭。利复星对HSA的能量转移效率为E=0.372, 利复星和人血清白蛋白的色氨酸残基的结合位置为R=1.933 nm。     

利福平与人血清白蛋白作用的荧光光谱

蛋白质是一类重要的生物大分子,它不仅是构成细胞内原生质的主要成分,也是生命现象的物质基础。对于蛋白质的探索是最复杂的课题之一。 以荧光光谱为手段,文章研究了药物利福平(Rifampicin capsules)与人血清白蛋白(HSA)的作用,测量发现利福平和人血清白蛋白的色氨酸残基的结合位置为R=2.567 nm, 临界距离R₀为2.433 nm。利福平-HSA的Lineweaver-Burk猝灭曲线的离解常数K_d=19.42×10-6 mol·L-1且结合常数K_s=5.149×104 L·mol-1。     

Separate and simultaneous binding effects of aspirin and amlodipine to human serum albumin based on fluorescence spectroscopic and molecular modeling characterizations: A mechanistic insight for determining usage drugs doses

The binding of aspirin (ASA) and amlodipine (AML) to human serum albumin (HSA) in aqueous solution was investigated by multiple techniques such as fluorescence quenching, resonance light scattering (RLS), three-dimensional fluorescence spectroscopy, FT-IR and zeta-potential measurements in an aqueous solution at pH=7.4. For the protein-ligand association reaction, fluorescence measurements can give important clues as to the binding of ligands to proteins, e.g., the binding mechanism, binding mode, binding constants, binding sites, etc. Fluorescence spectroscopy showed that ASA and AML could quench the HSA fluorescence spectra, and this quenching effect became more significant when both ASA and AML coexisted. The results pointed at the interaction between HSA and both drugs as ternary systems decreasing the binding constant and binding stability of the HSA-drug complex as a binary system. Therefore, by reducing the amount of drugs transported to their targets, the free drug concentration of the target would be reduced, lowering the efficacy of the drugs. It was demonstrated that there exists antagonistic behavior between the two drugs when it comes to binding of HSA. Furthermore, the fluorescence results also showed that the quenching mechanism of HSA-drug complexes as binary and ternary systems is a static procedure. The number of binding sites of HSA-ASA, (HSA-AML)ASA, HSA-AML and (HSA-ASA) AML were 1.31, 0.92, 1 and 0.93, respectively. Due to the existence of the antagonistic action between ASA and AML, the binding distance r was reduced. The results of synchronous fluorescence and three-dimensional fluorescence spectra showed that the antagonistic action between ASA and AML would alter the micro-environment around Trp and Tyr residues. Moreover, the simultaneous presence of ASA and AML during binding to HSA should be taken into account in multidrug therapy, as it induces the necessity of a monitoring therapy owing to the possible increase of uncontrolled toxic effects. Molecular dynamic studies showed that the affinity of each of the drugs to HSA was reduced in the presence of significant amounts of the other. In the interaction of HSA with both drugs, the zeta potential of the ternary system is more negative than its binary counterpart. The zeta-potential results suggested induced conformational changes on HSA that confirmed the experimental and theoretical results.     

On the structural formation of albumin

A microstructural analysis of the human serum albumin (HSA) samples dehydrated on a glass substrate from solutions with different concentrations of protein has been performed. The structuring effect of salt (NaCl) on the variations in the morphology of HSA interfaces has been shown experimentally. The substantiation of some structural effects has been given by taking into consideration the supramolecular organization and the polyelectrolyte nature of the protein molecule.