IPET:测定独个生物大分子三维空间结构的实验方法

蛋白质的动态特性和结构活性对于蛋白质功能的调控具有根本意义。传统的结构确定方法(包括X射线和电子显微镜单颗粒分析技术等)往往需要成千上万不同蛋白质分子的平均信号,因此难以确定蛋白质分子的动态结构。而电子显微断层成像技术是一种对独个生物个体结构从不同的观测角度照相、并计算来恢复该个体的三维结构密度图的方法。传统的冷冻电子断层成像重构方法采用整个大尺寸电镜图像进行重构计算,通常用来研究细菌、细胞切片等大尺寸生物个体在较低分辨率下的结构;由于分辨率的限制,不足以获得小尺寸的蛋白质分子的结构细节。最近,任罡研究小组提出一种独个生物颗粒的电子显微断层成像方法(individual-particle electron tomography,IPET)。该方法通过减小图像尺寸(直至所选区域只包含单个蛋白质分子)的策略,运用提出的FETR(focused electron tomography reconstruction)算法来提高独个大分子重构的分辨率。此方法不需要初始模型和大量分子的平均信号,同时能够容忍一定的测角误差。本文综述了IPET/FETR方法在确定独个分子结构过程中的具体步骤以及如何应用该方法来研究蛋白动态特性和结构变化特征。期望通过该综述和国内同行交流,分享最新的前沿研究,为赶超世界科技前沿的建设添砖加瓦。     

Microsensor Chip Integrated with Gold Nanoparticles‐Modified Ultramicroelectrode Array for Improved Electroanalytical Measurement of Copper Ions

This paper presents a microsensor chip integrated with a gold nanoparticles‐modified ultramicroelectrode array (UMEA) as the working electrode for the detection of copper ions in water. The microsensor chip was fabricated with Micro‐Electromechanical System technique. Gold nanoparticles were electrodeposited onto the surface of UMEA at a constant potential of ?0.3 V. The ratio d/R_b of interelectrode spacing (d) over the individual electrode’s radius (R_b) was investigated to improve the electrochemical performance. The UMEA with a d/R_b of 20 showed the best hemispherical diffusion mode, resulted in fast response time and high current response. The gold nanoparticles increased the active surface area of UMEA by not changing the geometries of UMEA, and the current response was increased further. Incorporating the optimized characteristic of UMEA and gold nanoparticles, the microsensor showed a good linear range from 0.5 to 200 µg L^?1 of copper ions in the acetate buffer solutions with the method of square wave stripping voltammetry. Compared with the gold nanoparticles‐modified disk electrode, the gold nanoparticles‐modified UMEA showed higher sensitivity (0.024 µA mm^?2 µg^?1 L) and lower limit of detection (0.2 µg L^?1). Water samples from river water and tap water were analyzed by the microsensor chip with recovery ranging from 100.7 % to 107.8 %.     

An efficient strategy for the extraction and purification of lignans from Schisandra chinensis by a combination of supercritical fluid extraction and high‐speed counter‐current chromatography

An efficient strategy for extracting and separating five lignans from Schisandra chinensis (Turcz.) Baill has been developed using supercritical fluid extraction (SFE) and high‐speed counter‐current chromatography (HSCCC) in the present study. First, the extraction was performed by a preparative SFE system under 15 MPa of pressure at 36°C for 4 h. Then, the SFE extract was successfully separated and purified by HSCCC with a two‐phase solvent system composed of n‐hexane/ethyl acetate/methanol/water (6:4:5:5, 6:4:6:4, 6:4:8:2, v/v) in a stepwise elution mode. The fractions were analyzed by HPLC, and the chemical structures of the products were identified by ESI‐MS and ¹H NMR spectroscopy. As a result, a total of 12.5 mg of schisandrin at 98.0% purity, 7.1 mg of gomisin A at 98.1% purity, 1.8 mg of schisantherin B at 93.3% purity, 4.4 mg of deoxyschisandrin at 92.9% purity, and 6.8 mg of γ‐schisandrin at 89.1% purity were obtained from 300 mg crude extract in a one‐step purification.     

Enantioseparation of chiral aromatic acids by multiple dual mode counter‐current chromatography using hydroxypropyl‐β‐cyclodextrin as chiral selector

This work concentrates on extending the utilization of multiple dual mode (MDM) counter‐current chromatography in chiral separations. Two aromatic acids, 2‐(6‐methoxy‐2‐naphthyl)propionic acid (NAP) and 2‐phenylpropionic acid (2‐PPA), were enantioseparated by MDM counter‐current chromatography using hydroxypropyl‐β‐cyclodextrin (HP‐β‐CD) as chiral selector. The two‐phase solvent systems consisting of n‐hexane/ethyl acetate 0.1 mol/L phosphate buffer pH 2.67 containing 0.1 mol/L HP‐β‐CD (7.5:2.5:10 for NAP and 7:3:10 for 2‐PPA, v/v/v) were used. Conventional MDM and modified MDM were compared according to peak resolution under current separation mechanism. The influence of elution time after the first‐phase inversion and number of cycles for MDM were investigated. Peak resolution of NAP and 2‐PPA increased from 0.62 to 1.05 and 0.72 to 0.84, respectively, using optimized MDM conditions. Being an alternative elution method for counter‐current chromatography, MDM elution greatly improved peak resolution in chiral separations.     

Efficient Method for Synthesis of the Derivatives of 5‐Arylidene Barbituric Acid Catalyzed by Aminosulfonic Acid With Grinding

Aminosulfonic acid is an environmentally friendly catalyst. Grinding a mixture of aromatic aldehydes, barbituric acid, and H₂NSO₃H at room temperature (without any solvent) gave 5‐arylidene barbituric acid in high yields, providing a simple and efficient route to synthesis of these compounds.     

Clean,One‐Pot Synthesis of Naphthopyran Derivatives in Aqueous Media

A general and practical one‐pot synthesis of naphthopyran derivatives using hexadecyltrimethylammonium bromide (HTMAB) as catalyst (10 mol%) is described. This method provides several advantages such as neutral conditions, high yields and simple workup procedure. The catalyst is low cost, facile, active, environmentally friendly, and reusable. In addition, water is chosen as a green solvent.     

Cascade reaction for 3-pyrrolines and pyrroles from nitroallylic acetates and N-mesyl 2-aminoethanones

Cascade reactions of nitroallylic acetates with methanesulfonyl 2-aminoethanones affords either the 3-pyrrolines or the pyrroles in one pot depending on reaction solvents and temperature. A possible mechanism for the entire sequence is proposed.     

Microstructural evolution during oxidative ablation in air for polyacrylonitrile based carbon fibers with different graphite degrees

Polyacrylonitrile‐based carbon fibers with different graphite degrees were oxidative ablated at 500 and 600 °C in air. By Thermal gravimetric (TG), Raman spectroscopy, X‐ray diffraction, and SEM, the mass loss, microstructure, and surface morphology of carbon fibers were investigated. The mass loss of carbon fiber increases linearly with increasing oxidative ablated time under 500 and 600 °C. The carbon fiber with higher graphite degree shows higher oxidative resistance, and the surface roughness increases gradually because of chemical ablation during the whole oxidation. A gloss morphology appears on the surface primarily because of physical denudation for carbon fibers with lower graphite degree and then burn off according to carbon and oxygen reaction. The crystallite size (La) decreases significantly, while interlayer spacing(d₀₀₂) remains nearly unchanged. SEM observation suggests the two kinds of ablation mechanisms for carbon fibers with different graphite degrees indicating that CC band in sp³ hybridization prefers to be attacked by oxygen molecule more than that in sp² hybridization during oxidation ablation in air. Copyright © 2012 John Wiley & Sons, Ltd.     

Synthesis,Crystal Structures,and Magnetic Properties of Two Copper(II) Radical Heterospin Complexes

Two heterospin complexes [Cu(NIT3Py)(cda)H₂O] · H₂O ( 1 ) and [Cu(NIT2Py)(cda)H₂O] · H₂O · CH₃OH ( 2 ) with Cu^II ions and pyridyl‐substituted nitronyl nitroxide radicals (NITxPy = 2‐(x′‐pyridyl)‐4,4,5,5‐tetramethyl‐imidazoline‐1‐oxyl‐3‐oxide, x = 3, 2; H₂cda = 4‐hydroxy‐pyridine‐2,6‐dicarboxylic acid) were synthesized and characterized structurally and magnetically. The single crystal structures show that the two complexes are both two‐spin complexes, in which the different radicals make the two complexes have different hydrogen bonding interactions to form 2D and 1D supramolecular network for complexes 1 and 2 , respectively. The magnetic measurements indicate that complexes 1 and 2 both exhibit antiferromagnetic interactions between Cu^II and radicals.