重组人超氧化物歧化酶化学修饰的初步研究

在高效表达重组人铜锌超氧化物歧化酶(rh Cu/Zn SOD),并纯化得到比活大于4000单位的 rh Cu/Zn SOD 纯品的基础上,采用活化酯法将聚乙二醇(PEG)与 rhCu/Zn SOD 交联,获得分子量约6万的 PEG-SOD 交联物.经 PEG 修饰的酶稳定性增强,表现为对酸、碱和热的耐受力均较未交联酶高.修饰酶的生物半衰期为15h,是天然酶的90倍,酶活性保留80%以上.还实验观察了修饰剂用量与修饰酶保留活性之间的关系.     

聚乙二醇修饰超氧化物歧化酶稳定性变化的研究

目的：了解各种分子量的聚乙二醇修饰超氧化物歧化酶(SOD)对其稳定的影响。方法：采用氰尿酰氯的修饰方法,用分子量为2000～20000的聚乙二醇(PEG)修饰SOD,测定SOD的酶活残存率及对修饰后的SOD的耐热、耐酸、耐碱和抗酶解能力进行研究。结果：聚乙二醇修饰后的SOD的耐热性、耐酸、耐碱以及抗酶解能力都明显增强,其中发现分子量6000的PEG修饰的SOD活件残余率较高,对酸、碱和酶的抗性较强。结论：分子量为6000PEG修饰的SOD的稳定性较高。     

长半衰期重组人超氧化物歧化酶的研制及性质

一种双亲有机化合物聚苯乙烯马来酸丁酯(SMA)经酰胺键与重组人铜锌超氧化物歧化酶(rhCu/Zn SOD)共价交联,制得修饰酶.当42%游离氨基被修饰时,保留酶活力为88%.酶蛋白主链结构在修饰前后变化不大.与天然酶相比,修饰酶的生物半衰期延长了22倍,抗蛋白水解酶能力亦有所增强.     

微生物谷氨酰胺转氨酶催化细胞色素c赖氨酸残基的定点修饰

研究微生物谷氨酰胺转氨酶(mTG)催化细胞色素c(Cytc)的PEG定点修饰的可行性,并优化修饰条件,研究PEG修饰对Cytc性质的影响。将单甲氧基聚乙二醇氨(mPEG-NH_2)与N-苄氧羰基-谷氨酰胺-甘氨酸(CBZ-QG)共价结合制备含谷氨酰胺残基的甲氧基聚乙二醇衍生物(N-苄氧羰基-谷氨酰胺-甘氨酰-单甲氧基聚乙二醇,CBZ-QG-mPEG);mTG分别催化mPEG-NH_2、CBZQG-mPEG(mTG)修饰Cytc,研究酶法定点修饰Cytc残基的可行性;改变酶的用量、温度、反应时间和p H等反应条件优化谷胺酰胺转氨酶催化修饰Cytc的条件。研究结果表明:(1)mPEG-NH_2不能作为mTG的底物修饰Cytc,甲氧基聚乙二醇氨(mPEG-NH_2)分子上引入谷氨酰胺残基后,在mTG的催化作用下了实现Cytc的PEG修饰,而且基于mTG的底物特异性实现了PEG定点修饰Cytc的赖氨酸(Lys)残基;(2)37℃温度下,p H 8.0的溶液中,1mg/ml的mTG催化修饰反应2h是最佳修饰反应条件;(3)化学法PEG修饰Cytc产物复杂,是多种多点修饰产物的混合物,酶法催化PEG修饰Cytc只产生单一产物;(4)与天然Cytc相比,修饰后Cytc的活力、稳定性都有所提高。提出的谷胺酰胺转胺酶催化修饰法解决了蛋白质Lys残基难以定点修饰的难题,拓展了mTG在蛋白质修饰方面的应用。     

棕榈酰化超氧化物歧化酶的制备及性质研究

为了增强超氧化物歧化酶的稳定性,用棕榈酸对其进行了修饰,在修饰条件下,酶分子表面氨基修饰率为55％时,酶的活力回收为63％。修饰后的酶在耐热、耐酸、耐碱、抗有机溶剂变性和抗蛋白水解能力上均高于天然超氧化物歧化酶,为将超氧化物歧化酶作成实用药物和进一步扩大其应用范围创造了条件。     

溶液介电常数对铜锌超氧化物歧化酶活性的影响

溶液介电常数对天然酶和修饰酶的活性影响不同,天然酶随介电常数增加而酶活性下降,修饰酶则反之,这表明静电相互作用在铜锌超氧化物歧化酶(Cu·Zn-SOD)与超氧阴离子(-O_2~(·-))反应过程中起着重要作用,酶分子活性中心附近ε-NH_3~+为O_2~(·-)进入活性中心提供静电吸引力。在有机溶剂中,SOD的构象会发生变化,从而导致酶活性降低。实验还表明,Cl~-对SOD有明显的抑制作用。     

活性氧所致超氧化物歧化酶疏水性的变化

用抗坏血酸-Fe(Ⅲ)和过氧化氢分别作用于铜锌超氧化物歧化酶(SOD),经疏水层析分离得到亲水型和疏水型SOD.用胰蛋白酶和胃蛋白酶分别作用于天然SOD,亲水型SOD及疏水型SOD,结果表明疏水型SOD较亲水型SOD及天然SOD易被降解,提示活性氧氧化修饰后的SOD对蛋白水解酶敏感性提高与其疏水性增高有关.     

月桂酸修饰超氧化物歧化酶的制备及其性质研究

用月桂酸对超氧化物歧化酶进行化学修饰以改善其稳定性.活化的月桂酸与SOD在40℃碱性条件下反应1h,再用Sephacryl S-200柱进一步纯化.修饰SOD活性回收率93%,比活为6000U / mg.具有较强的稳定性,基本上消除了免疫原性并明显地延长了在血液中的半衰期;该产品适用于化妆品和食品.     

静电作用与修饰铜锌超氧化物歧化酶的稳定性

铜锌超氧化物歧化酶(Cu, Zn-SOD)表面的赖氨酸经化学修饰后, 酶的稳定性显著提高. 赖氨酸被修饰后, 酶的电荷结构遂发生变化, 从而影响到酶分子电场. 使用FDPB方法(有限差分法求解Poission-Boltzman方程)计算了酶修饰前后的静电场变化, 以及对维持酶的结构稳定起重要作用的Cu, Zn配位结构的影响.结果表明, Cu, Zn配位体的两级离解常数在酶修饰后分别约下降10³, 10⁶.     

超氧化物歧化酶在反胶束体系中稳定性的研究

目的：考察反胶束体系组成对超氧化物歧化酶(SOD)抗氧化活性的影响。方法：以大豆磷脂氧化产生的氧自由基量为检测指标,考察温度、有机试剂以及表面活性剂浓度对反胶束体系中超氧化物歧化酶抗氧化活性的影响。结果：60℃时,反胶束中超氧化物歧化酶的抑制率由在水溶液中的52．48%升到75．38%,即SOD包封于反胶束中后热稳定性明显提高。表面活性剂可有效阻止有机溶剂对SOD的变性作用,SOD是不具界面活性的酶。结论：超氧化物歧化酶在反胶束体系中稳定性及抗氧化活性增强。     

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.     

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.     

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.

     

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).