核糖体蛋白L11(ribosome protein L11)是一种高度保守的蛋白质,是蛋白质合成过程中所必需的。L11由N-末端和C-末端两个结构域组成。L11的N-末端在蛋白质合成中作为分子开关,在多肽链的延伸阶段与延伸因子EF-G相互作用,对EF-G依赖的迁移...核糖体蛋白L11(ribosome protein L11)是一种高度保守的蛋白质,是蛋白质合成过程中所必需的。L11由N-末端和C-末端两个结构域组成。L11的N-末端在蛋白质合成中作为分子开关,在多肽链的延伸阶段与延伸因子EF-G相互作用,对EF-G依赖的迁移过程是必需的;在肽链终止阶段与肽链释放因子RF1相互作用,对RF1识别终止密码子UAG的功能是必需的。L11上有一个与噻唑类(thiazole)抗生素结合的靶位点,这种结合会抑制依赖延伸因子的核糖体的活性。展开更多
核糖体蛋白L11(ribosome protein L11)是一种高度保守的蛋白质.为研究真核生物的核糖体蛋白L11的功能,从八肋游仆虫(Euplotes octocarinatus)大核基因组中克隆到核糖体蛋白L11基因,构建了重组表达质粒pGEX-6p1-L11,通过谷胱甘肽-Sepharo...核糖体蛋白L11(ribosome protein L11)是一种高度保守的蛋白质.为研究真核生物的核糖体蛋白L11的功能,从八肋游仆虫(Euplotes octocarinatus)大核基因组中克隆到核糖体蛋白L11基因,构建了重组表达质粒pGEX-6p1-L11,通过谷胱甘肽-Sepharose 4B亲和层析,纯化了重组融合蛋白GST-L11.Pull down分析显示,八肋游仆虫的核糖体蛋白L11与第一类肽链释放因子eRF1a可以在体外相互作用.这一结果提示,与原核生物一样,低等真核生物的核糖体蛋白L11在肽链终止过程中可能起一定的作用.展开更多
A high-expression system of L11 was constructed and investigated its interaction with other elements of the ribosome using physicochemical methods. The gene rplK, coding for the protein L11 from the E. coli 50S riboso...A high-expression system of L11 was constructed and investigated its interaction with other elements of the ribosome using physicochemical methods. The gene rplK, coding for the protein L11 from the E. coli 50S ribosomal subunit was amplifyied, cloned and over-expressed. The protein L11 was purified under native and denaturing conditions, refolded and the structure of both proteins was compared. The protein L11 properly refolded from 6M urea after dialysis. Experiments on binding of proteins L11, RRF and EF-G from Escherichia coli were performed by ana-lytical centrifugation and Biacore. Specific binding between protein L11 and RRF by analytical cen-trifugation was not detected probably due to struc-tural reasons. These findings may be helpful in the design of new antibiotics that specifically disrupt the interactions in the “GTP-associated site” of the bac-terial ribosome, as many of them are not effective anymore. A common intrinsically disordered region of protein L11 was found to be the amino acid se-quence 86-97, while the residues 67-74, containing the linker region, are predicted to be disordered by DisEMBL.展开更多
文摘核糖体蛋白L11(ribosome protein L11)是一种高度保守的蛋白质,是蛋白质合成过程中所必需的。L11由N-末端和C-末端两个结构域组成。L11的N-末端在蛋白质合成中作为分子开关,在多肽链的延伸阶段与延伸因子EF-G相互作用,对EF-G依赖的迁移过程是必需的;在肽链终止阶段与肽链释放因子RF1相互作用,对RF1识别终止密码子UAG的功能是必需的。L11上有一个与噻唑类(thiazole)抗生素结合的靶位点,这种结合会抑制依赖延伸因子的核糖体的活性。
文摘A high-expression system of L11 was constructed and investigated its interaction with other elements of the ribosome using physicochemical methods. The gene rplK, coding for the protein L11 from the E. coli 50S ribosomal subunit was amplifyied, cloned and over-expressed. The protein L11 was purified under native and denaturing conditions, refolded and the structure of both proteins was compared. The protein L11 properly refolded from 6M urea after dialysis. Experiments on binding of proteins L11, RRF and EF-G from Escherichia coli were performed by ana-lytical centrifugation and Biacore. Specific binding between protein L11 and RRF by analytical cen-trifugation was not detected probably due to struc-tural reasons. These findings may be helpful in the design of new antibiotics that specifically disrupt the interactions in the “GTP-associated site” of the bac-terial ribosome, as many of them are not effective anymore. A common intrinsically disordered region of protein L11 was found to be the amino acid se-quence 86-97, while the residues 67-74, containing the linker region, are predicted to be disordered by DisEMBL.
