富硒螺旋藻中含硒藻蓝蛋白的纯化、结晶及初步晶体学研究

从富硒螺旋藻中提取含硒藻蓝蛋白,经凝胶色谱和离子交换色谱纯化,应用悬滴气相扩散法,采用(NH4)2SO4和PEG4000作沉淀剂,获得了该蛋白质晶体的两种晶型.晶型Ⅰ属于单斜晶系,晶胞参数a=10.80 nm,b=11.70 nm,c=18.40 nm,β=90.2°,晶体空间群属于P21.单位晶胞中每个晶体学不对称单位含12个(αβ)单体,晶体衍射的最高分辨率达0.28 nm.晶型Ⅱ为六方晶系,晶胞参数为a=b=15.5 nm,c=4.03 nm,晶体空间群属于P63.晶体衍射的最高分辨率达0.28 nm.单位晶胞中每个晶体学不对称单位含1个(αβ)单体.对分子在晶体中的可能堆积方式进行了讨论.     

杂质对蛋白质晶体影响研究进展

综述了杂质对蛋白质晶体生长影响研究领域的进展情况. 对可能的杂质来源以及杂质对结晶过程的影响进行了介绍.重点介绍了和结晶蛋白质分子结构相似的杂质分子的影响, 包括晶体成核、生长形态、表面形貌、生长动力学、质量等,以及杂质在晶体中的重新分配.     

蛋白质大分子的结晶

近几年研究蛋白质晶体的生长机理及过程日益受到重视 ,并逐步发展为蛋白质晶体学中的另一个重要的科学分支。1 .蛋白质结晶的基本过程蛋白质属于生物大分子 ,但其结晶的一般过程与盐、有机小分子等的结晶过程一样可分为 3个阶段进行 :( 1 )形成稳定的晶核 ;( 2 )由晶核生长为晶体 ;( 3)生长结束。1 .1　成核　　是指溶解在溶液中的分子或非晶态的低聚体 (二聚体 ,三聚体等 )以过饱和度为驱动力聚合在一起形成热力学稳态的点阵结构。Ｄｒｅｎｔｈ等[1] 研究发现 ,在蛋白质结晶后其溶液体系的自由能降低了 1 2～ 2 4ｋＪ/ｍｏｌ。另外 ,结晶蛋…     

杜仲抗真菌肽对黄曲霉的抑制作用(英文)

用纸片琼脂扩散法,研究了杜仲抗真菌肽EAFP3对黄曲霉(Aspergillus flavus)的抗真菌活性.研究结果表明,EAFP3对黄曲霉有明显抑制作用,最小抑菌浓度为125 μg/mL.     

多聚亚基蛋白质在固-液界面团聚的测量与表征

以血红蛋白和乙肝表面抗原(HBsAg)为模型,联用双偏振光干涉分析仪(DPI)和原子力显微镜(AFM)研究在固-液界面上的吸附行为.结果发现:当溶液环境中血红蛋白的质量浓度为1.08 mg/mL时,测得在硅羟基表面吸附的平均厚度为3.54 nm,蛋白颗粒直径达60～ 150 nm,远高于血红蛋白的分子尺寸,说明血红蛋白在固-液界面上发生团聚.当HBsAg质量浓度为0.67 mg/mL时,测得在硅羟基表面及DEAE琼脂糖表面吸附的平均厚度分别为1.06和23.69 nm,表面吸附的蛋白颗粒直径分别为288～401和520 ～ 971 nm,是单分子HBsAg尺寸的13.1～44.1倍,表明在模型蛋白表面都形成了大的团聚体.     

基于光纤式动态光散射的人血清白蛋白研究

人体尿液中血清白蛋白急剧增加会导致肾脏病发生几率增大,利用动态光散射技术(dynamic light scattering,DLS)研究人血清白蛋白有助于推动诊断肾脏病的早期发现。分析了人血清白蛋白的物理模型;利用单模光纤搭建了动态光散射实验系统,并配制了实验所需的人血清白蛋白水溶液;最后使用该系统研究了人血清白蛋白分子的扩散系数在不同蛋白浓度和pH值条件下的扩散系数。实验和分析结果表明库仑力对蛋白质的扩散起主要作用,在等电位点下(pH=5.2)库仑力的影响消失,蛋白质的扩散系数最小;在等电位点测量出扩散系数随浓度的增加而线性减小;在浓度5 mg/mL～40 mg/mL内互扩散系数Dm=D0[1-(0.00194±0.00008)],D0=(6.74±0.01)×10-7cm2/s为外推至零浓度下23℃时蛋白质的扩散系数。这里C为蛋白质浓度,实验测得人血清白蛋白的半径为(3.44±0.01)nm。     

N、P营养盐胁迫对两株布朗葡萄藻生长的影响

采用批次培养方法,在光照强度60、110mol/m2s下分别设置了7个不同的氮、磷浓度(N:0-3500g/L,P:15-775g/L),研究两株布朗葡萄藻(Botryococcus braunii)对氮、磷胁迫的敏感性差异,筛选高营养利用效率的优良藻株。结果表明:两株藻对氮磷营养胁迫的耐受性存在差异,B.braunii764株对氮胁迫具有较高耐受性,而B.braunii765株对磷胁迫具有较高耐受性。光照强度110mol/m2s,不同氮浓度下B.braunii764株其平均生长速率均显著高于其他各处理组;不同磷浓度下B.braunii765株其平均生长速率显著高于B.braunii764株。在试验设定的光照强度条件下,适当增加光照强度能够显著降低氮胁迫对布朗葡萄藻生长的抑制效应。在光照强度110mol/m2s下,氮浓度3500g/L时两株布朗葡萄藻平均生长速率与在正常Chu-10培养基条件下无显著差异。磷浓度775g/L时两株布朗葡萄藻的平均生长速率均显著低于正常Chu-10培养基条件,增加光照强度对磷胁迫下藻细胞的生长无显著作用。两株布朗葡萄藻在第2天时磷吸收与初始磷浓度呈正相关关系,氮吸收在3500g/L时出现饱和现象。布朗葡萄藻的生长更容易受到培养基中磷营养胁迫的影响。       

超滤膜分离纯化珊瑚藻溴过氧化物酶

利用超滤膜对珊瑚藻中溴过氧化物酶(EC 1.11.1.18,分子质量740kDa)进行分离纯化,对膜的截留分子质量、操作压力、起始蛋白质浓度、搅拌速率、pH、离子强度等条件进行了优化。超滤分离纯化最优条件为:截留分子质量为100kDa的聚偏氟乙烯(PVDF)膜,操作压力为0.02MPa,搅拌速率为600r/min,起始蛋白质浓度为0.1g/L,pH=6.0。对粗酶液先进行热沉淀纯化,再进行超滤纯化,溴过氧化物酶被纯化了21倍,比活为212U/mg,酶活回收率为96%。     

氮磷营养因子对赤潮异弯藻生长的影响

研究了N、P营养浓度对赤潮异弯藻(Heterosigma akashiwo)生长的影响.结果表明,该藻的生长速率与N、P营养因子浓度的关系符合Monod公式.在NO₃^--N浓度达到7.5 mg·L^-1时,赤潮异弯藻开始生长;浓度为3.75～75 mg·L^-1时,赤潮异弯藻的比生长速率与NO₃^--N浓度成正比关系.N营养充足时,赤潮异弯藻的最大生长速率μ_m-n=0.3475·d^-1,K_s-n=18.91 mg·L^-1.PO₄^--P浓度为0～1.0 mg·L^-1时,赤潮异弯藻的比生长速率与P浓度成正比关系;P营养充足时,赤潮异弯藻的最大生长速率μ_m-p=0.3024·d^-1,K_s-p=0.4086 mg·L^-1.N/P达到25后藻细胞浓度达到最大,表明N/P为25时最适合赤潮异弯藻生长.赤潮异弯藻最适合在N 37.5～225.0 mg·L^-1、P 5.0～50.0 mg·L^-1、N/P=25条件下生长.     

极端嗜酸热古菌Acidianus manzaensis膜蛋白提取方法的建立及应用

以万座嗜酸两面菌(Acidianus manzaensis)为研究对象,探索并优化其膜蛋白提取方法,以优化后的方法提取该菌分别以单质硫(S0)和亚铁(Fe2+)为能源底物进行生长时的膜蛋白质,并进行聚丙烯酰胺凝胶电泳(SDS-PAGE)研究膜蛋白在两种能源底物培养下的表达差异。首先通过80℃水浴60-70 min对A.manzaensis胞外黏附蛋白进行初步分离。其次比较了不同提取剂(Triton X-110,SDS和Triton X-114)对膜蛋白的提取效果,结果表明Triton X-114的提取效果较好,其最佳浓度为10%(w/v);比较了不同沉淀剂(三氯乙酸(TCA),丙酮,三氯乙酸/丙酮,甲醇和乙醇)对膜蛋白质沉淀的效果,结果表明TCA/丙酮的沉淀效果最不理想,会导致沉淀后低分子量蛋白发生缺失,而其他几种没有明显差别,综合比较选择较常用的丙酮作为膜蛋白提取的沉淀剂。最后基于优化后膜蛋白提取方法,分别对S0和Fe2+培养的A.manzaensis膜蛋白质进行提取及SDS-PAGE,结果发现分子量为35.6 k D和16.9 k D的蛋白只在A.manzaensis以S0生长时出现,表明这些蛋白质很可能在A.manzaensis硫氧化中发挥了重要作用;分子量为72 k D和26 k D的蛋白质在A.manzaensis以Fe2+生长时比其以S0生长时显著表达上调,表明此蛋白可能与A.manzaensis铁氧化相关。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Synthesis and antimicrobial studies of hydrazone derivatives of 4-[3-(2,4-difluorophenyl)-4-formyl-1H-pyrazol-1-yl]benzoic acid and 4-[3-(3,4-difluorophenyl)-4-formyl-1H-pyrazol-1-yl]benzoic acid

Microbial resistance to antibiotics is an unresolved global concern, which needs urgent and coordinated action. One of the guidelines of the Centers for Disease Control and Preventions (CDC) to combat antibiotic resistance is the development of new antibiotics to treat drug-resistant bacteria. In our effort to find new antibiotics, we report the synthesis and antimicrobial studies of 30 new pyrazole derivatives. These novel molecules have been synthesized by using readily available starting materials and benign reaction conditions. Some of these molecules have shown activity with MIC values as low as 0.78?µg/mL against four bacterial strains; Staphylococcus aureus, methicillin-resistant S. aureus, Bacillus subtilis, and Acinetobacter baumannii. Furthermore, active molecules are non-toxic to mammalian cell line.

     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Novel compounds with potent CDK9 inhibitory activity for the treatment of myeloma

Cyclin-dependent kinases (CDKs) and Polo-like kinases (PLKs) play key role in the regulation of the cell cycle. The aim of our study was originally the further development of our recently discovered polo-like kinase 1 (PLK1) inhibitors. A series of new 2,4-disubstituted pyrimidine derivatives were synthesized around the original hit, but their PLK1 inhibitory activity was very poor. However the novel compounds showed nanomolar CDK9 inhibitory activity and very good antiproliferative effect on multiple myeloma cell lines (RPMI-8226).