检索规则说明:AND代表“并且”;OR代表“或者”;NOT代表“不包含”;(注意必须大写,运算符两边需空一格)
检 索 范 例 :范例一: (K=图书馆学 OR K=情报学) AND A=范并思 范例二:J=计算机应用与软件 AND (U=C++ OR U=Basic) NOT M=Visual
作 者:Juanjuan Yang Jing Hong Ling Luo Ke Liu Chun Meng Zhi-liang Ji Donghai Lin
机构地区:[1]Institute of Pharmaceutical Biotechnology and Engineering,College of Biological Science and Biotechnology,Fuzhou University,Fuzhou 350108,China [2]State Key Laboratory of Stress Cell Biology,School of Life Sciences,Xiamen University,Xiamen 361102,China [3]High-Field NMR Center,Key Laboratory for Chemical Biology of Fujian Province,College of Chemistry and Chemical Engineering,Xiamen University,Xiamen 361005,China
出 处:《Acta Biochimica et Biophysica Sinica》2019年第2期139-149,共11页生物化学与生物物理学报(英文版)
基 金:the grants from the National Key Research and Development Project of China(No.2016YFA0500600);the National Natural Science Foundation of China(Nos.31700656 and 31670741);the Natural Science Foundation of Fujian Province(Nos.2017J05050 and 2017J01631).
摘 要:Mycobacterium tuberculosis(Mtb)is the key devastating bacterial pathogen responsible for tuberculosis.Increasing emergence of multi-drug-resistant,extensively drug-resistant,and rifampicin/isoniazid-resistant strains of Mtb makes the discovery of validated drug targets an urgent priority.As a vital translational component of the protein biosynthesis system,elongation factor Tu(EF-Tu)is an important molecular switch responsible for selection and binding of the cognate aminoacyl-tRNA to the acceptor site on the ribosome.In addition,EF-Tu from Mtb(MtbEF-Tu)is involved in the initial step of trans-translation which is an effective system for rescuing the stalled ribosomes from non-stop translation complexes under stress conditions.Given its crucial role in protein biosynthesis,EF-Tu is identified as an excellent molecular target for drug design.Here,we reported the recombinant expression,purification,biophysical characterization,and structural modeling of the MtbEF-Tu protein.Our results demonstrated that prokaryotic expression plasmids of pET28a-MtbEF-Tu could be expressed efficiently in Escherichia coli.We successfully purified the 6× His-tagged proteins with a yield of 16.8 mg from 1 l of Luria Bertani medium.Dynamic light scattering experiments showed that MtbEF-Tu existed in a monomeric form,and circular dichroism experiments indicated that MtbEF-Tu was well structured.Moreover,isothermal titration calorimetry experiments displayed that the purified MtbEF-Tu protein possessed intermediate binding affinities for guanosine-5′-triphosphate(GTP)and GDP.The GTP/GDP-binding sites were predicted by flexible molecular docking approach which reveals that GTP/GDP binds to MtbEF-Tu mainly through hydrogen bonds.Our work lays the essential basis for further structural and functional studies of MtbEF-Tu as well as MtbEF-Tu-related novel drug developments.
关 键 词:tuberculosis MtbEF-Tu PROTEIN biosynthesis PROTEIN expression and purification protein-guanine NUCLEOTIDE interaction
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在链接到云南高校图书馆文献保障联盟下载...
云南高校图书馆联盟文献共享服务平台 版权所有©
您的IP:216.73.216.249