基于共振瑞利散射法的人血清蛋白浓度的研究

文章针对人标准血清总蛋白浓度的定量测定, 建立了以四羧基金属酞菁锌为光谱探针检测蛋白质的共振光散射的新方法。该方法的实质是将人标准血清总蛋白和酞菁锌溶液混合, 当光源照射到样品池中的混合液时, 将激发出强烈的共振瑞利散射荧光信号, 根据信号强度的强弱, 计算血清蛋白浓度的大小。自行设计的测试装置采用了薄壁试管的样品皿, 减小了激光光波穿过溶液时的光程, 可得到更强的探测信号。测试结果表明: 根据数字示波器显示的探测器信号, 可得出电压的峰峰值, 体现了荧光的大小, 从而可以进一步计算出血清蛋白的浓度。     

细胞内蛋白质磁共振

细胞内的环境异常复杂,其中生物大分子的浓度超过300 mg/mL并占据着大约30%的细胞内空间. 然而,直到目前为止大多数的蛋白质研究仍然在细胞外或是甚至非常稀的缓冲液中进行. 细胞内磁共振（In-cell NMR）提供了一个可以直接非损伤地获取细胞内原子水平信息的方法. 虽然In-cell NMR在近10年里有了较大的发展,但是这一技术尚未成熟. 在该篇综述中,作者首先总结制约这一技术发展的因素以及研究中需要注意的事项,最后讨论了未来的发展方向.     

谷物种子中蛋白质含量测定的核反应分析法

分析了在高蛋白质谷物育种中对筛选方法的要求, 对谷物中蛋白质测定的方法进行了综述评价, 重点介绍了用于谷物蛋白质测定的核反应分析法的原理及特点. The requirements of the screening method for high protein content in grain breeding are analysed. Methods for the determination of protein content in grain seed are reviewed.The principle and the advantages of the nuclear reaction method for the measurement of protein content in grain seed are discussed in detail.     

双肽、多肽物质和蛋白质的紫外共振拉曼光谱研究现状

本文介绍了紫外共振拉曼光谱技术在双肽、多肽物质和蛋白质二级结构研究的现状和进展，双肽物质ＮＭＡ和ＧＬＹ—ＧＬＹ水溶液光异构化过程的紫外共振拉曼光谱研究，简述了紫外共振拉曼光谱实验技术要点。     

Two- and Three-Dimensional H/C PISEMA Experiments and Their Application to Backbone and Side Chain Sites of Amino Acids and Peptides

Two-dimensional ¹H/¹³C polarization inversion spin exchange at the magic angle experiments were applied to single crystal samples of amino acids to demonstrate their potential utility on oriented samples of peptides and proteins. High resolution is achieved and structural information obtained on backbone and side chain sites from these spectra. A triple-resonance experiment that correlates the ¹H–¹³C_α dipolar coupling frequency with the chemical shift frequencies of the α-carbon, as well as the directly bonded amide ¹⁵N site, is also demonstrated. In this experiment the large ¹H–¹³C_α heteronuclear dipolar interaction provides an independent frequency dimension that significantly improves the resolution among overlapping ¹³C resonances of oriented polypeptides, while simultaneously providing measurements of the ¹³C_α chemical shift, ¹H–¹³C dipolar coupling, and ¹⁵N chemical shift frequencies and angular restraints for backbone structure determination.     

Sensitivity-enhanced sim-CT HMQC PFG-HBHA(CO)NH and PFG-CBCA(CO)NH triple-resonance experiments

Short transverse relaxation times of Calpha and Cbeta single-quantum coherences reduce the sensitivity of triple-resonance experiments involving transfers of Calpha/Cbeta or Halpha/Hbeta coherences. Multiple-quantum line-narrowing techniques improve the relaxation properties of 13C coherences, thereby increasing the sensitivity of the experiment. In the present work, we describe PFG-CBCA(CO)NH and PFG-HBHA(CO)NH experiments that utilize heteronuclear multiple-quantum coherences in a simultaneous constant-time period to obtain completely decoupled spectra with improved sensitivity. Results indicate that approximately 30% of cross peaks show an average enhancement of approximately 15% in the CBCA(CO)NH experiment. In the related HBHA(CO)NH experiment, approximately 97% of the cross peaks show an average enhancement of approximately 40%.     

Superslow Backbone Protein Dynamics as Studied by 1D Solid-State MAS Exchange NMR Spectroscopy

Superslow backbone dynamics of the protein barstar and the polypeptide polyglycine was studied by means of a solid-state MAS 1D exchange NMR method (time-reverse ODESSA) that can detect reorientation of nuclei carrying anisotropic chemical shift tensors. Experiments were performed on carbonyl 13C in polyglycine (natural abundance) and backbone 15N nuclei in uniformly 15N-enriched barstar within a wide range of temperatures in dry and wet powders for both samples. Two exchange processes were observed in the experiments: molecular reorientation and spin diffusion. Experimental conditions that are necessary to separate these two processes are discussed on a quantitative level. It was revealed that the wet protein undergoes molecular motion in the millisecond range of correlation times, whereas in dry protein and polyglycine molecular reorientations could not be detected. The correlation time of the motion in the wet barstar at room temperature is 50-100 ms; the activation energy is about 80 kJ/mol. Previously, protein motions with such a long correlation time could be observed only by methods detecting chemical exchange in solution (e.g., hydrogen exchange). The application of solid-state MAS exchange spectroscopy provides new opportunities in studying slow biomolecular dynamics that is important for the biological function of proteins.     

Modifying the physicochemical properties,solubility and foaming capacity of milk proteins by ultrasound-assisted alkaline pH-shifting treatment

This study investigated the effects of different treatment of alkaline pH-shifting on milk protein concentrate (MPC), micellar casein concentrate (MCC) and whey protein isolate (WPI) assisted by the same ultrasound conditions, including changes in the physicochemical properties, solubility and foaming capacity. The solubility of milk proteins had a significant increase with gradual enhancement of ultrasound-assisted alkaline pH-shifting (p < 0.05), especially for MCC up to 99.50 %. Also, treatment made a significant decline in the particle size of MPC and MCC, as well as the turbidity of the proteins (p < 0.05). The foaming capacity of MPC, MCC, and WPI was all improved, especially at pH 11, and at this pH, the milk protein also showed the highest surface hydrophobicity. The best foaming capacity at pH 11 was the result of the combined effect of particle size, potential, protein conformation, solubility, and surface hydrophobicity. In conclusion, ultrasound-assisted pH-shifting treatment was found to be effective in improving the physicochemical properties and solubility and foaming capacity of milk proteins, especially MCC, with promising application prospect in food industry.     

Structure and stability characterization of pea protein isolate-xylan conjugate-stabilized nanoemulsions prepared using ultrasound homogenization

Preparation of pea protein isolate-xylan (PPI-X) conjugate-stabilized nanoemulsions using ultrasonic homogenization and the corresponding structure and environmental stability were investigated in this study. Conditions used to prepare nanoemulsions were optimized using a response surface methodology as follows: protein concentration 8.86 mg/mL, ultrasound amplitudes 57 % (370.5 W), and ultrasound time 16 min. PPI-X conjugate-stabilized nanoemulsions formed under these conditions exhibited less mean droplet size (189.4 ± 0.45 nm), more uniform droplet distribution, greater absolute value of zeta-potential (44.8 ± 0.22 mV), and higher protein adsorption content compared with PPI-stabilized nanoemulsions. PPI-X conjugate-stabilized nanoemulsions also exhibited even particle distribution and dense network structure, which might be reasons for the observed high interfacial protein adsorption content of conjugate-stabilized nanoemulsions. Moreover, better stability against environmental stresses, such as thermal treatment, freeze–thaw treatment, ionic strength and type, and storage time was also observed for the conjugate-stabilized nanoemulsions, indicating that this type of nanoemulsions possess a potential to endure harsh food processing conditions. Therefore, results provide a novel approach for the preparation of protein-polysaccharide conjugate-stabilized nanoemulsions to be applied as novel ingredients to meet special requirements of processed foods.     

Hardness and slip systems of orthorhombic hen egg-white lysozyme crystals

Hardness and slip systems by an indentation method were investigated on different habit planes of orthorhombic hen egg-white lysozyme (O-HEWL) crystals containing water. A dependence of the hardness on the water-evaporation time exhibits three stages as incubation, transition and saturated ones, as tetragonal (T)-HEWL crystals reported previously. The hardness values of (1 1 0), (0 1 0) and (0 1 1) habit planes of O-HEWL in the incubation stage or wet condition exhibits 6, 8 and 10 MPa, respectively. The hardness depends on indented planes but it is independent of the air-humidity and crystal volumes. These values correspond to the intrinsic hardness for O-HEWL crystals containing water. In the incubation stage, the slip traces are clearly observed around the indentation mark and the corresponding six kinds of slip systems are identified to be {0 1 1}<1 0 0>, {1 1 0}<1 1 0>, {0 1 1}<0 1 1>, {1 1 0}<0 0 1>, {1 0 0}<0 0 1> and {0 1 0}<0 0 1>.