A rat brain protein expression map including cytosolic and enriched mitochondrial and microsomal fractions

Proteomics is a powerful tool to screen brain protein expression but the methodology is hampered by low abundance of proteins or compartmentalization or overload of high-abundance proteins. It was therefore the aim of the study to determine the expression of brain proteins by using enriched cellular subfractions and pre-electrophoretic chromatographical separation of brain homogenates. We used two-dimensional electrophoresis with subsequent matrix-assisted laser desorption/ionization (MALDI) detection and characterization of brain proteins. Subfractionation into cytosolic, mitochondrial and microsomal compartments was performed by ultracentrifugation. Pre-electrophoretic fractionation of the cytosolic fractions was carried out by ion exchange column chromatography. We detected and identified a large series of 437 proteins in rat brain and have shown proteins specific for the individual subcellular compartments. These proteins included housekeeping, signaling, cytoskeletal, intermediary metabolism, antioxidant proteins on the one and neuron and synaptosomal specific proteins on the other hand. Using fractionations of brain homogenates we were able to improve the power of the method on forming the basis for brain protein expressional studies and providing a reference map as a powerful tool for the neuroscientist.     

Protein secondary structure and stability determined by combining exoproteolysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

In the post-genomic era, several projects focused on the massive experimental resolution of the three-dimensional structures of all the proteins of different organisms have been initiated. Simultaneously, significant progress has been made in the ab initio prediction of protein three-dimensional structure. One of the keys to the success of such a prediction is the use of local information (i.e. secondary structure). Here we describe a new limited proteolysis methodology, based on the use of unspecific exoproteases coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), to map quickly secondary structure elements of a protein from both ends, the N- and C-termini. We show that the proteolytic patterns (mass spectra series) obtained can be interpreted in the light of the conformation and local stability of the analyzed proteins, a direct correlation being observed between the predicted and the experimentally derived protein secondary structure. Further, this methodology can be easily applied to check rapidly the folding state of a protein and characterize mutational effects on protein conformation and stability. Moreover, given global stability information, this methodology allows one to locate the protein regions of increased or decreased conformational stability. All of this can be done with a small fraction of the amount of protein required by most of the other methods for conformational analysis. Thus limited exoproteolysis, together with MALDI-TOF MS, can be a useful tool to achieve quickly the elucidation of protein structure and stability.     

Investigation of ion-pair precipitates of selected alkoxylates and complex salts of specific metal cations by liquid secondary ion mass spectrometry

Liquid secondary ion (LSI) mass spectra of ion-pair precipitates obtained for Triton X-100 with strontium, lead, cadmium and mercury tetraphenylborates and for selected butoxylene-ethoxylene monoalkyl ethers with barium tetraiodobismuthate(III) are discussed. On the basis of LSI mass spectra, recorded in both positive and negative modes, the formulae of the ion-pair precipitates were determined. On the basis of B/E mass spectra, the fragmentation routes of [M - H + Ba](+) ions for butoxylene-ethoxylene monoalkyl ether complexes of barium and [M - H + Cd](+) ions for the Triton X-100 complex of cadmium are proposed.     

硫化氢对活性炭脱除单质汞的影响

H₂S是煤气中的含硫成分,活性炭能否催化氧化煤气中的H₂S形成活性硫,以促进H₂S和Hg⁰的协同脱除是值得研究的。 本文首先通过程序升温脱附和热力学分析了低温情况下H₂S对活性炭脱除Hg⁰的影响。 在较低吸附温度130 ℃下,H₂S不仅不能作为硫源,还使活性炭对Hg⁰的吸附显著减弱,这主要是由于H₂S对活性炭表面的吸附氧的消耗以及H₂S对含氧官能团的氧的取代反应造成的。 在此基础上,进一步研究了高温下H₂S对活性炭的影响,高温也不能使活性炭有效渗入单质硫。 所以无论低温还是高温情况下,利用H₂S作为硫源使Hg⁰以HgS的形式脱除并不是活性炭脱汞的可行手段。 本文揭示了较宽的温度范围内H₂S对单纯的活性炭脱除Hg⁰的影响机理,能为用于煤气脱汞的活性炭的设计以及作为负载时机理研究提供支持作用。     

Benchmarking the accuracy of coverage-dependent models: adsorption and desorption of benzene on Pt (1 1 1) and Pt3Sn (1 1 1) from first principles

Bimetallic catalysts have demonstrated properties favorable for upgrading biofuel through catalytic hydrodeoxygenation. However, the design and optimization of such bimetallic catalysts requires the ability to construct accurate, predictive models of these systems. To generate a model that predicts the kinetic behavior of benzene adsorbed on Pt (1 1 1) and a Pt₃Sn (1 1 1) surface alloy (Pt₃Sn (1 1 1)), the adsorption of benzene was studied for a wide range of benzene coverages on both surfaces using density functional theory (DFT) calculations. The adsorption energy of benzene was found to correlate linearly with benzene coverage on Pt (1 1 1) and Pt₃Sn (1 1 1); both surfaces exhibited net repulsive lateral interactions. Through an analysis of the d-band properties of the metal surface, it was determined that the coverage dependence is a consequence of the electronic interactions between benzene and the surface. The linear coverage dependence of the adsorption energy allowed us to quantify the influence of the lateral interactions on the heat of adsorption and temperature programmed desorption (TPD) spectra using a mean-field model. A comparison of our simulated TPD to experiment showed that this mean-field model adequately reproduces the desorption behavior of benzene on Pt (1 1 1) and Pt₃Sn (1 1 1). In particular, the TPD correctly exhibits a broadening desorption peak as the initial coverage of benzene increases on Pt (1 1 1) and a low temperature desorption peak on Pt₃Sn (1 1 1). However, due to the sensitivity of the TPD peak temperature to the desorption energy, precise alignment of experimental and theoretical TPD spectra demands an accurate calculation of the adsorption energy. Therefore, an analysis of the effect of the exchange-correlation functional on TPD modeling is presented. Through this work, we show the necessity of incorporating lateral interactions into theoretical models in order to correctly predict experimental behavior.     

Monitoring dendrimer conformational transition using 19F and 1H2O NMR

The conformational transition of a fluorinated amphiphilic dendrimer is monitored by the ¹H signal from water, alongside the ¹⁹F signal from the dendrimer. High-field NMR data (chemical shift δ, self-diffusion coefficient D, longitudinal relaxation rate R₁, and transverse relaxation rate R₂) for both dendrimer (¹⁹F) and water (¹H) match each other in detecting the conformational transition. Among all parameters for both nuclei, the water proton transverse-relaxation rate R₂(¹H₂O) displays the highest relative scale of change upon conformational transition of the dendrimer. Hydrogen/deuterium-exchange mass spectrometry reveals that the compact form of the dendrimer has slower proton exchange with water than the extended form. This result suggests that the sensitivity of R₂(¹H₂O) toward dendrimer conformation originates, at least partially, from the difference in proton exchange efficiency between different dendrimer conformations. Finally, we also demonstrated that this conformational transition could be conveniently monitored using a low-field benchtop NMR spectrometer via R₂(¹H₂O). The ¹H₂O signal thus offers a simple way to monitor structural changes of macromolecules using benchtop time-domain NMR.     

Electrochemical and structural investigations of oxide films anodically formed on ruthenium-plated titanium electrodes in sulfuric acid

The electrochemical characteristics of ruthenium oxides, formed on Ru-plated Ti electrodes in 0.5 M H₂SO₄ by potential cycling with different CV upper potential limits (E _SU), were systematically compared. The repeated potential cycling between 0.2 and 0.75 V activated the formation/reduction of surface Ru oxides with hysteretic behavior. This application of repeated CVs also modified the ability of Ru deposits for hydrogen adsorption/desorption. An irreducible Ru oxide accumulated on the electrode at potentials more positive than ca. 0.95 V, whose capacitive characteristics are applicable for electrochemical supercapacitors. This irreducible oxide was composed of an aggregate consisting of Ru in various oxidation states, bridged oxygen, OH and water in a 3D-like structure with a relatively ordered and compact nature, from the X-ray photoelectron spectroscopic and voltammetric results. The surface reconstruction of the Ru deposits induced by the repeated potential cycling with E _SU≥0.75 V was clearly observed from the SEM photographs. From the X-ray diffraction patterns, all the anodically formed Ru oxides showed an amorphous nature.     

基体辅助激光解吸电离飞行时间质谱和热裂解色谱/质谱研究聚酯型聚氨酯链结构

聚酯型聚氨酯样品用基体辅助激光裂解离子化质谱(MALDI-TOF/MS)和热裂解色谱质谱(PYGC/MS)方法进行了鉴定,PYGV/MS方法可检出PU的一些组成单元的化学结构,但PYGC/MS中大多数的峰,无法从常规的标准化合物的质谱数据库中检索到,而MALDI-MS方法,可明确测出PU试样各种单元的组合、聚合度及PU长链的序列分布,质量数范围可达2300.     

氮在含氧Mo(100)面上的吸附与反应

 用HREELS, AES, LEED和TDS考察了氮在含氧Mo(100)上的吸附和热脱附. 120 K下氮在含氧Mo(100)上吸附时存在着N—N伸缩振动频率2150和1600 cm-1, 分别对应于线式(γ态)和侧位(α态)两种分子吸附态. 升温引起γ态氮的脱附和α态氮的解离. 其中γ态氮的脱附峰温位于155 K, 遵循一级脱附动力学; 由α态解离生成的N原子占据Mo(100)的四重空位(即β态), 并在高于1?150 K的温度重新化合形成氮而脱附. 120 K时,氮的吸附是无序的; 吸附了氮的表面经1100 K退火后生成了有序的c(2×2)-N表面结构.     

聚(对苯二甲酸-1,3-丙二酯)环状低聚物的合成、表征和开环聚合反应

探索了由对苯二甲酰氯和1,3-丙二醇在“假高稀”条件下合成聚(对苯二甲酸-1,3-丙二酯)(PTT)环状低聚物的可行性.通过柱色谱分离了环状低聚物和线形低聚物;用核磁共振、质谱和元素分析表征了产物的化学结构;用GPC和HPLC研究了不同大小环的分布,发现在本文实验条件下合成的PTT环状低聚物主要由二、三、四、五和七聚体构成,其中环状三聚体含量最多,没有发现环状六聚体的存在.PTT环状低聚物的熔程为92.3～222.6℃,熔融后是无色、透明的低粘度液体.于250℃将PTT环状低聚物分别在辛酸亚锡、1-乙基-3-氯四丁基锡氧烷、钛酸四丁酯和三氧化二锑催化下进行开环聚合反应,制备了特性粘数为0.18～0.49dL/g的聚合物.