Interaction of Globular Albumins with the Silica Surface

Bovine serum albumin macromolecules interact with vicinal hydroxyl groups of the silica surface at carbonyl groups and with isolated surface hydroxyl groups at the imide groups. Geminal surface hydroxyl groups behave as single adsorption sites relative to albumin adsorption. The concentration of such groups on a quartz surface was evaluated. The secondary structure of globular albumins is altered as the result of interaction in the silica–globular albumin system. Extended -segments appear on the surface and the concentration of -spirals decreases. This alteration leads to change in the tertiary globular structure of the albumin. The adsorption of albumin macromolecules on the silica surface is inclined.     

Interaction of Globular Plasma Proteins with Water‐Soluble CdSe Quantum Dots

The interactions between water‐soluble semiconductor quantum dots [hydrophilic 3‐mercaptopropionic acid (MPA)‐coated CdSe] and three globular plasma proteins, namely, bovine serum albumin (BSA), β‐lactoglobulin (β‐Lg) and human serum albumin (HSA), are investigated. Acidic residues of protein molecules form electrostatic interactions with these quantum dots (QDs). To determine the stoichiometry of proteins bound to QDs, we used dynamic light scattering (DLS) and zeta potential techniques. Fluorescence resonance energy transfer (FRET) experiments revealed energy transfer from tryptophan residues in the proteins to the QD particles. Quenching of the intrinsic fluorescence of protein molecules was noticed during this binding process (hierarchy HSA<β‐Lg <BSA, lower binding affinity for hydrophobic protein molecules). Upon binding with QD particles, the protein molecules underwent substantial conformational changes at the secondary‐structure level (50 % helicity lost), due to loss in hydration.     

Lateral Force Microscopy Study of the Friction between Silica Surfaces

The friction between a single silica glass sphere and a flat silica glass surface has been investigated using lateral force microscopy. Two types of silica surface were investigated, one fully hydroxylated and the other partially dehydroxlated. These samples have markedly different wetting properties and are known as hydrophilic and hydrophobic silica, respectively. The lateral friction force as a function of applied normal load has been collected for five different-sized spheres for both surface types. Analysis of the friction versus load data indicates that the hydrophilic surfaces have a molecularly smooth contact. For the hydrophobic surfaces, it is not clear whether the friction-load response is caused by a smooth or an asperity-dominated contact zone. Copyright 2000 Academic Press.     

CdS纳米晶与多肽分子相互作用研究

研究了半导体CdS纳米晶的表面功能化及荧光光谱特性, 并利用静电/配位自组装方法实现了多肽和CdS纳米晶的生物无机偶联, 研究了纳米晶多肽偶联体系的荧光光谱以及多肽与CdS纳米晶之间的相互作用. 结果表明: 含巯基多肽对CdS纳米晶表面形成完善包覆, 消除CdS纳米晶表面缺陷, 使CdS纳米晶荧光增强; 含端氨基多肽使CdS纳米晶荧光出现先升后降趋势; 其余不含巯基和氨基的多肽均猝灭CdS纳米晶荧光, 猝灭机制属于形成化合物所引起的静态猝灭, 它们的结合常数约为2×10⁴, 结合位点数约为0.87～1.00.     

Adhesion between Silica Particle and Mica Surfaces in Water and Electrolyte Solutions

An atomic force microscope (AFM) is used to study the adhesion between a silica sphere and a mica plate in pure water and solutions of monovalent cations (LiCl, NaCl, KCl, and CsCl). It is found that the adhesive force depends not only on the electrolyte concentration but also on the hydration enthalpy of cations and the contact time of the particle on the surface. Possible mechanisms by which the observed phenomena can be explained consistently are discussed extensively. It is suggested that the adhesive force is closely related to the structure of the layer of cations and water molecules adsorbed on the surfaces: the strong adhesive force is obtained when highly hydrated cations (Li(+), Na(+)) are adsorbed to form a thick but weakly adsorbed layer, while the weak adhesive force is observed when poorly hydrated cations (Cs(+), K(+)) are adsorbed to form a thin but strongly adsorbed layer. Copyright 2000 Academic Press.     

偶氮类染料与蛋白质作用的分光光度研究

本文研究了阴离子偶氮染料在酸性条件下与蛋白质的作用，在最佳反应条件下制定了标准工作曲线，讨论了蛋白质变剂的作用以摩尔比法测定了蛎结合数。     

Spectrophotometric Study on the Interaction between Arsenazo M and Proteins

Dye binding protein assays are commonly used in biochemical and clinical laboratories, and the reaction mechanism is still under investigation1-2. In this paper, the interaction of Arsenazo M with serum proteins was studied. Procedure In most experiments, 1.20 mL of Arsenazo M (5.0010-4 mol/L), 2.0 mL of C-L buffer solution, 3.0 mL alcohol and certain amount of protein standard solutions or samples were added to 10 mL volumetric flasks, then diluted to mark with water, and mixed thor…     

Resolving the Discrepancies Between Empirical and Rayleigh Charge Limiting Models for Globular Proteins

Starting with the Rayleigh charge limiting model, a slightly different approach is used to account for the well-known discrepancy that exists between the said model and experimental ESI MS data for globular proteins. It is shown using published datasets that for globular proteins, the mass density ρ exhibits a weak second-order dependence on its mass M, according to ρ(M)∝ M^-α, α ~ 0.14. A direct equivalence established between ESI MS and x-ray techniques suggests a minimum but critical surface tension of 15.6?±?5.2 mN/m for the droplet at the liquid-to-gas phase transition point. The packing density factor η for globular proteins is believed to lie between 1 (very tightly packed) and 4.6 (less tight, natively packed). While the Rayleigh charge limiting model has been linked historically to the CRM (J. Chem. Phys. 49:2240–2249, 1968; Anal. Chim. Acta 406:93–104, 2000), this paper does not expressly seek to justify the CRM, but rather uses empirical data and existing knowledge across subfields to help build a consistent picture of ESI MS phenomena that might be difficult to explain otherwise. These results would be useful in molecular dynamics (MD) simulations, understanding liquid-to-gas phase transitions and in opening up new routes for cross-calibration between ESI MS, IM MS, NMR and x-ray crystallography studies.

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Detailed Evidence for an Unparalleled Interaction Mode between Calmodulin and Orai Proteins

Calmodulin (CaM) binds most of its targets by wrapping around an amphipathic α‐helix. The N‐terminus of Orai proteins contains a conserved CaM‐binding segment but the binding mechanism has been only partially characterized. Here, microscale thermophoresis (MST), surface plasmon resonance (SPR), and atomic force microscopy (AFM) were employed to study the binding equilibria, the kinetics, and the single‐molecule interaction forces involved in the binding of CaM to the conserved helical segments of Orai1 and Orai3. The results consistently indicated stepwise binding of two separate target peptides to the two lobes of CaM. An unparalleled high affinity was found when two Orai peptides were dimerized or immobilized at high lateral density, thereby mimicking the close proximity of the N‐termini in native Orai oligomers. The analogous experiments with smooth muscle myosin light chain kinase (smMLCK) showed only the expected 1:1 binding, confirming the validity of our methods.     

Chronopotentiometric Analysis of Proteins at Charged Electrode Surfaces

Protein properties and functions are strongly dependent on the structure and amino acid content. In this work, catalytic hydrogen evolution reaction (CHER) of five proteins (human serum albumin, lysozyme, β‐synuclein, H2 A and H3 histones) were studied using constant current chronopotentiometric stripping (CPS) with the aim to find out the association between protein content and its electrochemical response. We have shown that the height and potential of CPS peak H in dependence on accumulation potential differed for the studied proteins, while the CPS peak area was almost the same for all of them. CV and CPS peaks H of Cys‐containing proteins appeared at less negative potentials in comparison to proteins without Cys, suggesting easier CHER. Acidic and basic proteins not containing Cys can be also recognized due to their different CPS response after their adsorption at the positive and negative charged interface.     

The Hydrophobicity Study of Silica and Glass Surfaces Modified with Polyalkylhydrosiloxanes

A procedure based on the analysis of the shape of a sessile drop was used to measure the contact angles of water in air, _a = 93°–97°, for quartz and glass plates modified with polyalkylhydrosiloxanes. The specific heat of wetting, q = 36 mJ/m², and the surface pressure of adsorbed water film, = 48 mJ/m², were determined from adsorption-calorimetric experiment on macroporous disperse silica modified with polymethylhydrosiloxane GKZh-94M. The contact angle in saturated water vapors, _v = 84°, was calculated from these values. It was shown that the liquid-phase modification of silica and glass with polyalkylhydrosiloxanes leads to distinct hydrophobization of the studied surfaces.     

Proteins Entrapped in Silica Monoliths Prepared from Glyceroxysilanes

The preparation of discrete polyol based silane precursors derived from glycerol by a simple one-pot process is described. These polyol-based silanes could be hydrolyzed under mild pH conditions and upon gelation resulted in the formation of optically clear, monolithic, mesoporous silica. The hydrolysis and condensation reactions lead to cure rates that are very sensitive to ionic strength, but are almost unaffected by pH in contrast to those of alkoxysilanes derived from primary alcohols such as Si(OEt)₄. Residual glycerol in the silica monolith could be removed by washing, or could be left in the silica to reduce the magnitude of shrinkage during long term storage. The biocompatible glyceroxysilane precursors lead to materials that were able to retain the activity of entrapped enzymes over repeated cycles of use for periods of up to several months.     

Analyzing the Interaction between Anthocyanins and Native or Heat-Treated Whey Proteins Using Infrared Spectroscopy

The color stability of anthocyanins (ACN) has been shown to be improved by interaction with whey proteins (WP). In this study, we explore the ACN–WP interaction using Fourier transform infrared spectroscopy (IR). ACN from purple corn, grape, and black carrot (50 μM) were evaluated. IR spectra (4000–700 cm⁻¹) were collected for native and preheated (40–80 °C) WP (5 mg/mL) and ACN–WP mixtures at pH 7.4. Soft independent modeling of class analogy was used to analyze the IR data. The WP secondary structure changed after heat treatments and after interaction with ACN. As expected, the WP α-helices decreased, and β-sheet increased after heat treatment. The intensities of the WP amide I and II bands decreased after ACN addition, revealing a decrease in the WP α-helix content. Higher preheating temperatures (70–80 °C) resulted in a more disordered WP structure that favored stronger WP–ACN interactions related to amide III changes. Addition of ACN stabilized WP structure due to heat denaturation, but different ACN structures had different binding affinities with WP. WP structure had less change after interaction with ACN with simpler structures. These results increase our understanding of ACN–WP interactions, providing a potential strategy to extend anthocyanin color stability by WP addition.     

利用原子力显微镜研究在单分子水平上DNA与组蛋白的相互作用

核小体是DNA被压缩成染色质的第一级压缩结构. 为了更深刻描述DNA与组蛋白的相互作用,利用透析方法将DNA和组蛋白构建成核小体,并且应用原子力显微镜进行单分子水平上的观察. 实验结果表明利用透析方法得到了核小体的“串珠型排列”,并且对在两种不同的结合条件下DNA与组蛋白的相互作用进行了观察和比较.     

Hydrophobic Interaction Chromatography of t-RNA's and Proteins

Abstract

Various microparticulate siliceous bonded stationary phases having weakly hydrophobic ligates were developed for HPLC of proteins and t-RNA's by hydrophobic interaction chromatography (HIC). It was confirmed that optimal separation of different types of biopolymers can be obtained by using a set of stationary phases having appropriate hydrophobic properties. Thus, the separation of t-RNA's is best carried out on stationary phases which are more hydrophobic than those optimal for HIC of proteins. Plots of log k' of both proteins and t-RNA's against the salt molality in the eluent yielded straight lines at sufficiently high salt concentrations in the eluent. The limiting slopes represent the hydrophobic interaction parameter for the particular chromatographic system and can serve as measures of the hydrophobic character of either the biopolymer or the stationary phase. Stationary phases with covalently bound polyether chains at the surface were found to be most suitable for HIC of proteins and t-RNA's.     

水化介孔氧化硅表面酸碱平衡常数的测定

采用自动电位酸碱滴定技术研究了水化介孔氧化硅的表面脱质子反应.基于悬浮液的酸碱滴定数据,用Gran图法计算得到了介孔氧化硅表面氢离子键位点浓度.利用FITEQL4.0软件,采用双电层恒电容模型计算得到了相应的表面酸碱平衡常数.结果表明,与无定形氧化硅不同,介孔氧化硅表面存在对应于双齿表面硅羟基≡Si2OH和单齿表面硅羟基≡SiOH的两个脱质子常数,分别为pKa1=6.78±0.15,pKa2=10.25±0.22.根据测定得出的介孔氧化硅表面脱质子常数,构建了不同pH条件下,介孔氧化硅表面优势组分分布图,并讨论了表面电容预赋值对氧化硅表面组分分布的影响.     

蛋白质与三氯乙酸相互作用的共振散射光谱研究及分析应用

三氯乙酸（TCA）与酪蛋白、明胶、γ－球蛋白、HSA、BSA结合形成缔合微粒，均使体系的共振散射信号显著增强，在470nm处产生一共振散射峰。在选定条件下，几种蛋白质在一定浓度范围内与I470nm呈线性关系。建立了定量测定蛋白质的共振散射光谱分析新方法。该法操作简便，灵敏度高，线性范围宽，重现性较好，可用于多种蛋白质的测定。本法应用于合成样品及人血清样品中蛋白质的定量分析，结果满意。     

Stereoselective Interaction between DNA and Stable Chiral Surfaces Modified with 1,2‐Diphenylethylenediamine Enantiomers

This work described an interesting phenomenon of the stereoselective adsorption behaviors of DNA on stable chiral surfaces which were modified with 1,2‐diphenylethylenediamine enantiomers on gold electrodes. The modification process and electrochemical characterization of the chiral surfaces were measured by cyclic voltammetry (CV). The stereoselective adsorption behaviors of DNA on the two chiral surfaces were investigated via atomic force microscopy (AFM), CV, electrochemical impedance spectroscopy (EIS) and quartz crystal microbalance (QCM). All results confirmed that (1R,2R)‐1,2‐diphenylethylenediamine modified surface had stronger interaction with DNA molecules than (1S,2S)‐1,2‐diphenylethylenediamine modified surface, and the chirality of the surfaces created an different effect on the morphology and adsorption quantity of DNA.     

Structures and Unexpected Dynamic Properties of Phosphine Oxides Adsorbed on Silica Surfaces

Solid‐state NMR spectroscopy of selected phosphine oxides adsorbed on silica surfaces establishes the surface mobilities, even of phosphine oxides with high melting points. Crystal structures of the adducts Ph₃PO ? HOSiPh₃ and Cy₃PO ? H₂O indicate that the interactions with silica involve hydrogen bonding of the P?O group to adsorbed water and surface silanol groups.     

某些氨羧络合型染料与蛋白质相互作用的共振瑞利散射研究

用共振瑞利散射(RRS)光谱法研究了酚酞络合剂、百里酚酞络合剂以及钙黄绿素等氨羧络合型染料与人血清白蛋白(HSA)、牛血清白蛋白(BSA)、α-糜蛋白(α-Chy)、卵白蛋白(Ova)、溶菌酶(Lys)等水溶性蛋白质的相互作用,发现只有作为氨羧络合剂咕吨染料的钙黄绿素与蛋白质反应能使RRS显著增强,并观察到特征的RRS光谱,而作为氨羧络合型三苯甲烷的酚酞络合剂、百里酚酞络合剂作用不明显.文中对于钙黄绿素与蛋白质的反应条件、影响因素、反应产物的光谱特征及有关分析化学性质进行了研究,并拟定了用钙黄绿素RRS测定水溶性蛋白质的方法.该方法有高的灵敏度(对不同蛋白质的检出限在4.0～40.0 ng·mL^-1之间)和较好的选择性,可用于人血清中蛋白质的测定.