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目的:利用N端缺失10个氨基酸的葡激酶(recombinant staphylokinase,rSaK)重组质粒,构建了表达可溶性rSaK126蛋白的工程菌,并研究不同条件下工程菌诱导表达目的蛋白含量的差异及纯化途径。 方法:采用细菌活化和培养方法诱导目的蛋白,并用SDS-PAGE测其含量,应用层析技术纯化蛋白。 结果:成功构建表达重组葡激酶的工程菌,表达的重组葡激酶蛋白约占菌体总蛋白的50%,经纯化后回收率为60%,纯度达99%以上。结论:成功构建高效表达重组葡激酶的工程菌,并获得了高含量、高纯度的目的蛋白。 相似文献
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本综述从葡激酶的溶栓作用机制,包括与纤溶酶(原)等因子的结合与作用,葡激酶的高级结构,抗原性问题等方面概括了近年来有关葡激酶作为亲一代溶栓剂的研究成果,并指出进一步利用蛋白质工程,对葡激酶进行分子改造的设想。 相似文献
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本综述从葡激酶的溶栓作用机制,包括与纤溶酶(原)等因子的结合与作用,葡激酶的高级结构,抗原性问题等方面概括了近年来有关葡激酶作为亲一代溶栓剂的研究成果,并指出进一步利用蛋白质工程,对葡激酶进行分子改造的设想。 相似文献
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采用阴离子交换和凝胶过滤色谱法纯化大肠杆菌高密度培养所表达的重组葡激酶-水蛭素融合蛋白(rSFH),经SDS-PAGE和RP-HPLC分析纯度达到98%以上,每升发酵液得率约0.7g。同时利用疏水色谱、MALDI-TOF对纯化过程中出现的rSFH同源二聚体的分析和表面疏水面积的计算及利用高效排阻色谱(HPSEC)分析NaCl、温度对rSFH的可逆二聚化行为的影响,认为疏水作用在rSFH的可逆二聚化行为中发挥着重要作用。 相似文献
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葡萄球菌激酶作为新型溶栓剂的研究进展 总被引:2,自引:0,他引:2
近年来对溶栓剂的研究取得了很好的成果,主要集中在单链尿激酶型纤溶酶原激活剂(scuPA),组织型纤溶酶原激活剂(tPA),葡萄球菌激酶(SaK)等。葡萄球菌激酶(SaK)是一种激活溶纤维蛋白的制剂,它与纤溶酶原(Plg)形成1∶1的复合体,使后者转变为纤溶酶(Pli)后激活其他分子变为Pli。从葡激酶的溶栓作用机制,包括与纤溶酶(原)等因子的结合作用,葡激酶的高级结构,抗原性等问题以及近年来有关葡激酶作为新一代溶栓的研究进展进行了综述,并指出进一步利用蛋白质工程,对葡激酶进行分子改造的设想。 相似文献
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同源模建人微小纤溶酶原(microplasminogen ,mplg)的三维结构,建立葡激酶(staphylokinase,Sak),mplg计算机分子对接的结构模型,模型与已有的实验结果基本相符。为研究葡激酶N端结构与功能的关系,进一步改造葡激酶分子,根据复合物结构模型设计了N端缺失15个氨基酸的葡激酶突变体。 相似文献
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[目的]为解决溶栓后再栓塞问题,构建N-端含RGD(Arg-Gly-Asp)序列的葡激酶双功能突变体.研究突变体的表达和纯化,并进行性质分析.[方法]将突变后的葡激酶突变体序列连入pBV220质粒,转化大肠杆菌BL21进行表达.阳离子交换、凝胶过滤和阴离子交换三步层析法纯化表达产物,采用溶圈法对纯化产物进行生物学活性测定,并测定纯化产物对血小板聚集的抑制效应.[结果]PAGE扫描结果显示,葡激酶突变体蛋白在大肠杆菌BL21中的表达量约占菌体蛋白总量的40%~50%;三步层析纯化后,HPLC测定其纯度可达95%.酪蛋白凝胶板溶圈法测得其比活性分别为10.8×104和11.0×104HU/mg,与野生型葡激酶活性相当;且具有明显的抗血小板聚集活性,血小板聚集仪测定其血小板聚集抑制率分别为10.72%和19.71%,明显高于野生型葡激酶血小板聚集抑制率.本实验利用pBV220载体高效表达了葡激酶突变体基因,得到了高纯度、高活性的突变体蛋白,为葡激酶生产产业化和临床应用奠定了良好的基础. 相似文献
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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. 相似文献
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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 相似文献
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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. 相似文献
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A.A. Zakeyah J. Whitt C. Duke D.F. Gilmore D.G. Meeker M.S. Smeltzer M.A. Alam 《Bioorganic & medicinal chemistry letters》2018,28(17):2914-2919
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. 相似文献
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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. 相似文献
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Zsófia Czudor Mária Balogh Péter Bánhegyi Sándor Boros Nóra Breza Judit Dobos Márk Fábián Zoltán Horváth Eszter Illyés Péter Markó Anna Sipos Csaba Szántai-Kis Bálint Szokol László Őrfi 《Bioorganic & medicinal chemistry letters》2018,28(4):769-773
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). 相似文献