title: Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine.
abstract: We demonstrate here that a high concentration (40-70%) of pyridine, an aromatic tertiary amine catalyst, is able to promote translation on ribosomes without the presence of soluble protein factors or chemical energy sources. Compared with Monro's fragment reaction [Methods Enzymol. 20, 472-481 (1971)] which reflects only the peptidyltransferase step, this novel translation system can produce polypeptides with chain lengths of at least several tens of residues depending on the template RNA. In the presence of 60% pyridine, poly(U) and poly(UC) promoted incorporation of the respective amino acids, phenylalanine and serine-leucine, twofold, whereas poly(A) promoted the incorporation of lysine by only 25%. The degrees of polymerization of phenylalanine and lysine were up to the decamer and around 40mer, respectively. In poly(UC)-dependent oligo(serine-leucine) synthesis, oligopeptides with a serine and leucine alternate sequence were the main products. This novel pyridine system evidently differs from the non-enzymatic translation system reported by Gavrilova and Spirin [FEBS Lett. 17, 324-326 (1971)]; the former system displays partial resistance toward deproteinization reagents such as SDS and proteinase K, whereas the latter system is completely sensitive.
authors.affiliation: Department of Chemistry and Biotechnology, Faculty of Engineering, University of Tokyo.
authors.forename: I
authors.initials: I
authors.lastname: Nitta
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.forename: T
authors.initials: T
authors.lastname: Nojima
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
mesh_terms: D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D011725:Pyridines / Q000378:metabolism*; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D013698:Templates, Genetic
chemical_list: D011725:Pyridines; C023666:pyridine