Analysis of Adsorbed Urea and Alkylthiourea on Alumina Surface by Inelastic Electron Tunneling Spectroscopy

Inelastic electron tunneling spectroscopy is used in the analysis of chemisorbed species of urea and alkylthiourea on the alumina surface using an Al-Al₂O₃-Pb tunnel junction. Tunneling spectra of urea, thiourea, N,N′-dimethylthiourea (DMTU), and tetramethylthiourea (TMTU) were taken and analyzed for designated band positions and intensities by comparing with the corresponding IR spectrum. These molecules seem to exhibit similar adsorption behavior onto alumina surface. Thiourea and urea interact with the surface through the nitrogen lone pairs and orient perpendicularly on the Lewis acid site of the alumina surface. DMTU and TMTU are still adsorbed through the nitrogen atoms, but the molecular plane has a somewhat inclined orientation to the alumina surface.     

聚乙烯醇微透镜阵列的制备及其固载细胞色素c的直接电化学

利用自组装法以聚苯乙烯有序多孔膜为膜板制备聚乙烯醇微透镜阵列膜.在pH6.89的磷酸盐(PBS)缓冲溶液中,固载在聚乙烯醇微透镜阵列膜修饰的玻碳电极上的细胞色素c于0.072V(vs·Ag/AgCl)处显示一对准可逆的氧化还原峰,是细胞色素c血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.考察了扫速、溶液pH及支持电解质浓度等因素对细胞色素c电子传递的影响,体系的表观异相电子传递速率常数k0为2.98×10-6cm·s-1.     

基于偏振光波导分光光谱技术原位研究纳米金-细胞色素c 自组装过程

时间分辨偏振光波导分光光谱技术是一种用于研究表面分子吸附动力学的强大工具. 利用该技术实时、原位监测纳米金与细胞色素c的静电自组装过程, 发现随着吸附层数的增加, 纳米金粒子吸附层产生的局域等离子体共振(LSPR)吸收峰发生了红移, 而且在横磁(TM)模式下的红移比横电(TE)模式下的红移更快; 细胞色素c在纳米金表面的吸附导致LSPR吸收峰的峰位和强度在TM模式下显著红移和升高, 相比之下, TE模式下的LSPR吸收峰无明显变化. 对实验数据的分析验证了纳米金在细胞色素c单分子层表面的吸附动力学行为遵循扩散控制模型, 细胞色素c在纳米金单粒子层表面的吸附动力学行为遵循Langmuir等温吸附模型, 进一步估算了细胞色素c在纳米金表面的吸附速率常数、脱附速率常数和吸附自由能.     

Acid Denaturation and Refolding of Cytochrome c on Silica Surface

Denaturation of oxidized cytochrome c (cyt c) adsorbed to a hydrophilic fused silica surface was studied by UV‐VIS attenuated total reflection (ATR) spectroscopy using a multiple optical pass system newly developed by this lab. Cyt c surface adsorption at neutral pH gave an adsorption equilibrium constant of K_a = 2 × 10⁵ M^?1 and a surface coverage at 63% of a monolayer saturation. Protein unfolding by acid denaturation was studied by equilibrating surface bound cyt c with acid buffers ranging in pH from 5 to 2. Protein orientation and surface coverage were calculated based on a theoretical model developed in previous work. The average heme tilt angle (44°) was found to be independent of pH, implicating protein‐surface interactions as the dominant factor governing adsorption. A non‐random molecular orientation distribution of cyt c on the surface was observed, providing further support for the dominance of protein‐surface interactions. It was shown that when denaturing acid buffers were removed and replaced with a neutral buffer cyt c refolded, assuming their original conformation. The combination of unique, yet applicable, science and laboratory skills involved in this project had a tremendous impact on the authors‘ undergraduate curriculum, making it ideal for capstone project development.     

Topological and Electron‐Transfer Properties of Yeast Cytochrome c Adsorbed on Bare Gold Electrodes

The redox metalloprotein yeast cytochrome c was directly self‐chemisorbed on “bare” gold electrodes through the free sulfur‐containing group Cys102. Topological, spectroscopic, and electron transfer properties of the immobilised molecules were investigated by in situ scanning probe microscopy and cyclic voltammetry. Atomic force and scanning tunnelling microscopy revealed individual protein molecules adsorbed on the gold substrate, with no evidence of aggregates. The adsorbed proteins appear to be firmly bound to gold and display dimensions in good agreement with crystallographic data. Cyclic voltammetric analysis showed that up to 84 % of the electrode surface is functionalised with electroactive proteins whose measured redox midpoint potential is in good agreement with the formal potential. Our results clearly indicate that this variant of cytochrome c is adsorbed on bare gold electrodes with preservation of morphological properties and redox functionality.     

The Transient Complex of Cytochrome c and Cytochrome c Peroxidase: Insights into the Encounter Complex from Multifrequency EPR and NMR Spectroscopy

We present a novel approach to study transient protein-protein complexes with standard, 9 GHz, and high-field, 95 GHz, electron paramagnetic resonance (EPR) and paramagnetic NMR at ambient temperatures and in solution. We apply it to the complex of yeast mitochondrial iso-1-cytochrome c (Cc) with cytochrome c peroxidase (CcP) with the spin label [1-oxyl-2,2,5,5-tetramethyl-Δ3-pyrroline-3-methyl)-methanethiosulfonate] attached at position 81 of Cc (SL−Cc). A dissociation constant K_D of 20±4×10⁻⁶ M (EPR and NMR) and an equal amount of stereo-specific and encounter complex (NMR) are found. The EPR spectrum of the fully bound complex reveals that the encounter complex has a significant population (60 %) that shares important features, such as the Cc-interaction surface, with the stereo-specific complex.     

Optical properties of Yeast Cytochrome c monolayer on gold: an in situ spectroscopic ellipsometry investigation

The adsorption of Yeast Cytochrome c (YCC) on well defined, flat gold substrates has been studied by Spectroscopic Ellipsometry (SE) in the 245-1000 nm wavelength range. The investigation has been performed in aqueous ambient at room temperature, focusing on monolayer-thick films. In situ δΨ and δΔ difference spectra have shown reproducibly well-defined features related to molecular optical absorptions typical of the so-called heme group. The data have been reproduced quantitatively by a simple isotropic optical model, accounting for the molecular absorption spectrum and film-substrate interface effects. The simulations allowed a reliable estimate of the film thickness and the determination of the position and the shape of the so-called Soret absorption peak that, within the experimental uncertainty, is the same found for molecules in liquid. These findings suggest that YCC preserves its native structure upon adsorption. The same optical model was able to reproduce also ex situ results on rinsed and dried samples, dominated by the spectral features associated to the polypeptide chain that tend to overwhelm the heme absorption features.     

双氰胺在金团簇上吸附的密度泛函理论和表面增强拉曼光谱研究

双氰胺是氰胺的二聚体,具有亚氨式和氨式两种互变异构体.将表面增强拉曼光谱(SERS)与密度泛函理论(DFT)结合,研究了互变异构的双氰胺分子在金表面的吸附行为.通过理论计算获得了亚氨式和氨式双氰胺分子的能量、分子轨道和光谱信息,以及双氰胺分子吸附在金簇表面的SERS响应.计算结果表明两种异构化的双氰胺分子都与Au₃簇形成较稳定的复合物,并且双氰胺分子中N₂原子优先吸附在金簇表面.拉曼实验结果与计算结果较为吻合,进一步说明具有互变异构的双氰胺分子在金基底中共存,并通过N₂原子垂直吸附到金表面,符合SERS电磁场增强机制.     

表面带负电荷氨基酸残基的突变对细胞色素b5与细胞色素c结合和识别的影响

利用核磁共振方法研究表面带不同负电荷氨基酸残基突变后的细胞色素b₅与细胞色素c的结合与识别.结果表明,静电作用在细胞色素b₅与细胞色素c的结合过程中有着重要的贡献,而且这些静电贡献在一定程度上具有累加性,E48的贡献略大于E44.同时还证明Browniandynamicssimulations优化出的Glu48-Lys13,Glu56-Lys₈₇,Asp60-Lys86和heme6-propionate-Tml₇₂(细胞色素b₅的残基排在前面)的结合方式在溶液中的确存在.细胞色素b₅突变体(E48,E56/A,D60/A)及[Cr(oxalate)₃]^3-对细胞色素c的表面结合竞争实验表明,细胞色素c表面结合区Site仍然同细胞色素b₅突变体(E48,E56/A,D60/A)有结合作用,只是结合强度上相对于野生细胞色素b₅同细胞色素c的结合有所降低.这表明除上述的Brownian dynamics simulations模型外,尚有其它如Salemme模型等的结合方式,这也揭示出细胞色素b₅和细胞色素c之间的结合是比较动态的.     

Synthesis, Characterization and Selective Metal Binding Properties of Physically Adsorbed 2-Thiouracil on the Surface of Porous Silica and Alumina

The physical adsorption of 2-hydroxy-2-mercaptopyrimidine (2-thiouracil) on the surface of porous silica gel and basic alumina is studied and described. Four silica and alumina materials were identified on the basis of surface coverage determination by thermal desorption method. Modified aluminas were found to be physically adsorbed with 2-thiouracil by 0.14, 0.54, and 0.43–1.63mmolg^–1. Modified silica materials (I–IV) were identified by adsorption of 2-thiouracil (0.344–0.446mmolg^–1). The metal sorption, binding and selectivity properties of these modified alumina and silica materials were studied and evaluated by a series of di- and trivalent metal ions. Aluminas (II–IV) were found to have high capacity in selective extraction of cadmium (II) from aqueous solutions compared to other tested metal ions. The highest metal capacity (1.0mmolg^–1) was found for cadmium (II) on modified aluminas (IV). Potential application of the modified aluminas (I–IV) for selective removal and extraction of 1.0ngmL^–1 of cadmium (II) from natural seawater samples was performed and compared with the modified silicas (I–IV). The modified aluminas (II–IV) showed a significantly higher capacity than the modified silicas for cadmium (II) with an average percentage recovery value of 97.8±0.4%. The matrix effect of seawater was found to play an insignificant role in the atomic absorption spectrophotometric analysis of cadmium (II).     

电化学和紫外-可见吸收光谱法研究pH和金纳米粒子对细胞色素c的影响

制备了粒径均匀、平均粒子尺度为(4.7±0.6)nm,表面修饰3-巯基丙酸(MPA)的金纳米粒子(AuNPs).利用电化学和紫外-可见(UV-Vis)吸收光谱研究了pH和AuNPs对细胞色素c(Cytc)结构的影响.UV-Vis吸收光谱结果表明,pH为7.5-3.0时,Cytc和Cytc-AuNPs复合物的结构没有发生明显变化.当pH=2.0时,Cytc和Cytc-AuNPs复合物的Soret谱峰位置均发生明显移动,说明pH诱导其构象发生变化.循环伏安(CV)结果表明,表面修饰了MPA的AuNPs能促进Cytc和电极之间的电子传输,与修饰了柠檬酸根的AuNPs相比,其生物兼容性更好.pH的变化会引起CV中Cytc峰电流的改变和峰电位的移动.随着pH值的降低,Cytc电活性的量逐渐减小,并且pH诱导Cytc发生不可逆变性.AuNPs的引入使自由态的Cytc耐酸性增强,而使得吸附态的Cytc耐酸能力减弱.     

基于磁球表面的抗原决定基印迹法用于选择性识别细胞色素c的研究

本工作合成了一种核壳型的抗原决定基磁性分子印迹聚合物,并用于选择性识别目标蛋白细胞色素c(Cytochrome c,Cyt c)。制备过程中先用溶剂热法合成Fe_3O_4磁性纳米粒子,然后加入Cyt c其C端的九肽作为模板,进行一段时间的预组装,最后加入多巴胺盐酸盐(DA)溶液,调节反应体系的p H使多巴胺聚合在磁球表面。洗脱掉模板后,即得到分子印迹聚合物。优化DA的用量使聚合物达到最佳的吸附效果。在最优条件下,制得的印迹聚合物对目标蛋白有较好的吸附选择性,并且有良好的重复利用性。此外,用抗原决定基做模板制得的聚合物的吸附容量和印迹因子明显优于用相应蛋白质做模板的情况。     

Influence of Light Scattering by Residual Alumina Nanoparticles on the Analysis of Surfactants Adsorption Using Spectroscopy

The adsorption of a surfactant mixture, based on an anionic surfactant, sodium dodecyl benzenesulfonate (SDBS) and a nonionic surfactant (Triton X-100, or TX100), on alumina nanoparticles was determined by solution depletion method combined with spectrometric measurement. It is shown that the light scattering, originated from the residual adsorbent alumina particles in the supernatant after centrifugation separation, interferes with the measurements of absorbance of the surfactant molecules, and therefore constitutes an error source for determination of the surfactant concentration in the supernatant by spectrometric means. The intensity of this light scattering, namely the influence of the residual alumina nanoparticles upon the surfactant adsorption, was related to the surfactant adsorption and its equilibrium concentration and varied among a batch. In this paper we report a Kalman filter method in order to eliminate the variational scattering background caused by non-separated residual alumina nanoparticles in each supernatant. This method is of interest as it is simple, easy to carry out and of high precision.     

TATB固体与表面吸附水的相互作用研究

TATB（1，3，5-三氨基-2，4，6-三硝基苯）是最著名的耐热钝感炸药；水在 TATB表面的吸附作用研究具有理论和实用双重意义。在B3LYP/6-31G~（**）水平上 ，在对TATB晶体（001）表面作周期性计算的基础上经基组叠加误差（BSSE）校正 ，求得TATB的表面能为-19.90 kJ·mol~(-1)，与实验值良好相符；首次求得水在 TATB（001）面的吸附能为-10.25kJ·mol~(-1)；重点讨论了吸附前后能带和电子 结构的变化。     

CrO3.CH3NH2.HCl／Al2O3的制备及其对苯偶姻体系的氧化反应

ＣｒＯ３·ＣＨ３ＮＨ２·ＨＣｌ／Ａｌ２Ｏ３的制备及其对苯偶姻体系的氧化反应张贵生＊石启增陈密峰蔡昆（河南师范大学化学系新乡４５３００２）关键词ＣｒＯ３·ＣＨ３ＮＨ２·ＨＣｌ／Ａｌ２Ｏ３，苯偶姻，苯偶酰，氧化１９９６－０８－１８收稿，１９９６－１１－０...     

Influence of Nanoscale Surface Topography and Chemistry on the Functional Behaviour of an Adsorbed Model Macromolecule

The functional behaviour of a model macromolecule (fibrinogen) adsorbed at the nanofabricated solid‐liquid interface was found to be strongly influenced by the local topographic structure of the interface. Protein molecules bound at topographically structured surfaces (either chemically homogeneous or heterogeneous 40‐nm diameter and 10‐nm deep pits) were found to bind platelets significantly faster than uncoated substrates whereas proteins bound to flat (R_a 1 nm) substrates were not. During the initial interaction, the chemistry of the underlying substrate apparently does not affect the macromolecules' functional behaviour.     

细胞色素c在单壁碳纳米管表面的固定、直接电子转移及电催化

应用吸附法将细胞色素c(Cytoc)固定在单壁碳纳米管(SWNT)表面.红外光谱(IR)显示被固定的Cytoc能保持原有的空间结构,没有发生变性.循环伏安测试表明,Cytoc在SWNT表面能发生稳定的直接电子转移,其i~E曲线上出现一对良好的、几乎对称的氧化还原峰.式量电位E0’基本不随扫速的增加而变化(在20 mV~120 mV/s的扫速范围内,E0’平均值为0.165±0.001V).实验同时给出,吸附在SWNT表面的Cytoc仍能保持其对H2O2电化学还原的生物电催化活性.     

细胞色素c／L-半胱氨酸修饰金电极检测亚硝酸根

采用循环伏安法、交流阻抗法研究了组装在L-半胱氨酸(L-Cys)修饰金电极上的细胞色素c(Cyt c)电化学行为;采用电化学方法以及紫外-可见光谱对电极进行表征.结果表明:通过静电吸附作用组装在L-Cys修饰金电极上的细胞色素c保持了良好的生物和电化学活性,用Cyt c/L-Cys修饰金电极检测亚硝酸根,响应电流与亚硝酸根浓度在5.0×10-6～4.5×10-4mol·L-1范围内呈线性关系,线性回归方程为Ip=0.031 c 8.165×10-6,相关系数为0.999 5,检出限(S/N=3)为1.5×10-7mol·L-1.电极用于模拟样品及咸菜样品中亚硝酸根的测定,回收率为89.0%～116.0%.     

Structures of Cytochrome b_5 Mutated at the Charged Surface-Residues and Their Interactions with Cytochrome c

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