Molecular recognition of DNA–protein complexes: A straightforward method combining scanning force and fluorescence microscopy

Combining scanning force and fluorescent microscopy allows simultaneous identification of labeled biomolecules and analysis of their nanometer level architectural arrangement. Fluorescent polystyrene nano-spheres were used as reliable objects for alignment of optical and topographic images. This allowed the precise localization of different fluorescence particles within complex molecular assemblies whose structure was mapped in nanometer detail topography. Our experiments reveal the versatility of this method for analysis of proteins and protein–DNA complexes.     

Yeast KEX2 protease and mannosyltransferase I are localized to distinct compartments of the secretory pathway

The KEX2 protease (product of the KEX2 gene) functions late in the secretory pathway of Saccharomyces cerevisiae by cleaving the polypeptide chains of prepro-killer toxin and prepro-alpha-factor at paired basic amino acid residues. The intracellular vesicles containing KEX2 protease sedimented in density gradients to a position distinct from those containing mannosyltransferase I (product of the MNN1 gene), a marker enzyme for the Golgi complex. The recovery of intact compartments containing these enzymes approached 80% after sedimentation. We propose that the KEX2 protease and mannosyltransferase I reside within distinct compartments.     

Protein Engineering Strategies for Improved Pharmacokinetics

Protein therapeutics have gained momentum in recent years and become a pillar in treating many diseases and the only choice in several ailments. Protein therapeutics are highly specific, tunable, and less toxic than conventional small drug molecules. However, reaping the full benefits of therapeutic proteins in the clinics is often hindered by issues of immunogenicity and short half-life due essentially to fast renal clearance and enzymatic degradation. Advances in polymer chemistry and protein engineering allowed overcoming some of these limitations. Strategies to prolong the half-life of proteins rely on increasing their size and stability and/or fusing them to endogenous proteins (albumin, Fc fragment of antibody) to hijack physiological pathways involved in protein recycling. On the downside, these modifications might alter therapeutic proteins structure and function. Therefore, a compromise between half-life and activity is sought. This review covers half-life extension strategies using natural and synthetic polymers as well as fusion to other proteins and sheds light on genetic engineering strategies and chemical and enzymatic reactions to achieve this goal. Promising strategies and successful applications in the clinics are highlighted.     

不同制备条件对SpA分子印迹聚合微球聚合效果的影响

本文用金黄色葡萄球菌蛋白A(SpA)作为模板,以丙烯酰胺为功能单体,利用反相悬浮聚合法合成了SpA分子印迹聚合微球.通过扫描电镜观测不同条件下聚合微球的聚合效果,研究不同制备条件对SpA分子印迹聚合微球聚合效果的影响,从而筛选制备聚合微球最佳工艺;并检测了制备的分子印迹聚合物对SpA的吸附性能.结果表明,反相悬浮聚合法合成SpA分子印迹微球的最佳工艺条件为:分散剂乙基纤维素(EC)加入量0.20 g/100 mL(甲苯);甲苯与水的比例10:3;交联度10%;引发剂加入方式为缓慢滴加;反应温度50℃.反相悬浮聚合法制备的SpA分子印迹聚合微球对SpA的吸附量:6.956×10-3μmol/g.     

BP神经网络结合变量选择方法在牛奶蛋白质含量检测中的应用北大核心CSCD

牛奶中的蛋白质含量会影响牛奶的品质,利用高光谱图像的光谱特征信息研究对牛奶蛋白质含量预测的可行性。本文提出一种基于竞争性自适应重加权算法(competitive adaptive reweighted sampling, CARS)和连续投影算法(successive projections algorithm, SPA)结合多层前馈神经网络(back propagation, BP)的预测建模方法,实验以含有不同浓度蛋白质的牛奶为对象,利用可见光/近红外高光谱成像系统共采集到5种牛奶共计250组高光谱数据,通过实验对比选择采用标准化方法对获取到的吸收光谱预处理,然后采用CARS结合SPA筛选特征波长,得到18个特征波长,建立CARS-SPA-BP模型,经过试验,CARS-SPA-BP模型的训练集决定系数和测试集决定系数R;和R;分别达到0.971和0.968,训练集均方根误差(root mean square error of calibration,RMSEC)和测试集均方根误差(root mean square error of prediction,RMSEP)达到了0.033和0.034。研究发现,采用CARS结合SPA筛选的牛奶特征波长建立的多层前馈神经网络模型,其模型预测结果与全波长建模相比并没有明显降低,因此将CARS结合SPA用于波长筛选并且结合BP神经网络基本可以完成对牛奶蛋白质含量的预测。为验证CARS-SPA-BP模型的预测能力,在相同数据环境下,使用较为传统的偏最小二乘回归(partial least squares regression, PLSR)进行建模,实验结果表明,CARS-SPA-BP相较于PLSR,R;和RMSEP均有明显提升。研究表明,CARS-SPA-BP可充分利用牛奶光谱特征信息实现较高精度的牛奶蛋白质含量检测。     

基于负染－电镜的Aβ体外聚集表征测试要点分析

本文应用电镜负染色技术,对阿尔茨海默病（ Alzheimer′s disease, AD）中所涉及的β淀粉样肽（β?amyloid peptide,Aβ）的聚集规律进行了分析。通过对样品的浓度、pH值、温度和时间等设置系列梯度,以及调整负染试剂的种类、浓度和染色时间,优化了Aβ寡聚体样品制备以及电镜观察的最佳条件,获得了清晰的Aβ纤维图像并用以估算纤维长度。实验结果表明：经过六氟异丙醇给予单体化处理,并溶于无水二甲基亚砜（ DMSO）的Aβ（100μmol·L－1）,在pH7?2的PBS中4℃孵育24 h,Aβ寡聚体形态清晰,且Aβ42寡聚体比Aβ40寡聚体聚集趋势更明显。在pH 8?0的硼酸缓冲液中,Aβ42（40μmol·L－1）在37℃孵育72 h后,纤维形成明显;而Aβ纤维样品经过煮沸后再制备负染样品,所得电镜图像更为清晰,便于对纤维长度和结构进一步分析。因此电镜负染色技术,可作为一种快捷,直观的Aβ体外聚集形貌表征的质控方法。     

ORIGIN OF A DOMINANT BEER PROTEIN IMMUNOCHEMICAL IDENTITY WITH A β-AMYLASE-ASSOCIATED PROTEIN FROM BARLEY

Two-dimensional immunoelectrophoretic techniques were used to elucidate the origin of a dominant beer antigen 1. Immunochemical identity was found between a protein Z associated with β-amylase in extracts of barley and one form of the beer antigen 1a. Immunochemical identity was also found between a papain-modified form of protein Z and the antigen form 1b from beer. The results indicate that protein Z is very resistant to degradation and denaturation during malting and brewing.     

髓鞘碱性蛋白对脂筏微区结构影响的模式化研究

本文主要通过原子力显微镜(AFM)和Langmuir单层膜技术研究不同浓度的髓鞘碱性蛋白(MBP)与"脂筏"(PC/SM/Cholesterol)模型相互作用形成仿生膜的物理特性.利用表面压缩模量(Cs-1)和二维维里状态方程对π-A曲线进行分析,计算了MBP与"脂筏"单层膜分子间相互作用的第二维里系数.根据二维维里状...     

芝麻蛋白发泡性、持水力的理论研究

本文比较系统地研究了芝麻蛋白的发泡性和持水力等功能特性，探讨了不同蛋白质浓度、ｐＨ值、离子强度和温度因素对芝麻蛋白发泡性和持水力的影响，进行了芝麻蛋白功能特性的变化规律的研究，从理论上解释了芝麻蛋白的发泡性以及持水力的变化特点，为芝麻蛋白在食品工业上的应用提供了依据。     

Accelerated Fermentation Process for the Manufacture of Fish Sauce Using Histidine

Addition of histidine accelerated hydrolysis of fish protein during fermentation in the manufacture of fish sauce and after 4 mo fermentation yielded a product typical of traditional fish sauce. The liquefaction rate of the histidine-treated sauce was faster than that of the control. The degree of hydrolysis was much greater in the histidine-added sauces than in the control. Most amino acids were higher in the histidine sauces compared to commercial patis sauce (as reference). Addition of histidine to the fish mixture during fermentation did not increase the histamine content of the sauces.