Stoichiometry of GTP hydrolysis in a poly(U)-dependent cell-free translation system: Determination of GTP/peptide bond ratios during codon-specific elongation and misreading

• Published in: FEBS letters Vol. 178; no. 2; pp. 283 - 287
• Main Authors: Gavrilova, L.P., Kakhniashvili, D.G., Smailov, S.K.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 10-12-1984; Elsevier
• Subjects: Biological and medical sciences; Cell-free translation system; Codon; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; GTP hydrolysis; Guanosine Triphosphate - metabolism; Hydrolysis; Kinetics; Matrix-bound poly (U) Functionally active ribosome; Misreading; Molecular and cellular biology; Molecular genetics; Peptide Biosynthesis; Peptide Chain Elongation, Translational; Peptides; Poly U; Protein Biosynthesis; Ribosomes - metabolism; Translation; Translation. Translation factors. Protein processing; Misreading; Translation; GTP hydrolysis; Cell-free translation system; Matrix-bound poly (U) Functionally active ribosome; Hydrolysis; Stoichiometry; Error; GTP; In vitro translation; Elongation
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(84)80617-1

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 Mg
2+ concentration (6 mM MgCl
2), 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).