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Stoichiometry of GTP hydrolysis in a poly(U)‐dependent cell‐free translation system

机译:依赖于Poly(U)的无细胞翻译系统中GTP水解的化学计量

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>The Stoichiometry of GTP hydrolysis during peptide elongation in the processes of codon-specific translation and misreading of polyuridylic acid was determined in a cell-free system in which all ribosomes were active in peptide synthesis. Ribosomes carrying oligophenylalanine presynthesized on poly(U) covalently bound to Sepharose were used. In the codon-specific translation of poly(Phe) on poly(U)-Sepharose at optimal Mg2+ concentration (6 mM MgCl2), the ratio of GTP cleaved to Phe polymerized was found to be about 2 (+- 0.1). Under the same conditions but during misreading (elongation of polyleucine on poly(U)-Sepharose) the GTP/Leu ratio increased 10 times (from 16 to 25 in different experiments).
机译:在无细胞系统中测定了密码子特异性翻译和多尿酸的误读过程中肽延伸过程中GTP水解的化学计量。在该系统中,所有核糖体均在肽合成中起作用。使用携带寡聚苯丙氨酸的核糖体预先合成在与Sepharose共价结合的poly(U)上。在最佳Mg 2 + 浓度(6 mM MgCl 2 )下,poly(U)-Sepharose上poly(Phe)的密码子特异性翻译中,GTP的比例被切割发现聚合的Phe的分子量约为2(±0.1)。在相同条件下但在误读期间(聚(U)-琼脂糖上的聚亮氨酸伸长),GTP / Leu比增加了10倍(在不同实验中从16增至25)。

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