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- W973404156 abstract "This chapter discusses the chemical characterization of the polypeptidyl-soluble RNA (sRNA) complex in Escherichia coli cell-free systems. The chemical characterization of a polypeptidyl-sRNA complex requires that both the polypeptide and sRNA moieties may be recognized and that the complex is purified from free sRNA, amino acyl-sRNA, and the other proteins. Although these requirements are not readily achieved, the use of synthetic polynucleotides as messenger RNA's (mRNA's) in a cell-free system may yield polypeptides with very unusual properties. Particularly, poly A and poly U direct the synthesis of polylysyl-sRNA and polyphenylalanyl-sRNA. In neither of these cases is the polypeptide released from sRNA, except for a small fraction released by spontaneous hydrolysis of the complex during the incubation. The chapter describes the method of detecting polyphenylalanyl-sRNA, where the reagents used include crude preincubated Escherichia coli extract, 32P-labeled sRNA, L-phenylalanine-14C, polyuridylic acid, and Sephadex G-200. In principle, this method can be used for rather large polypeptides (usually greater than 20 amino acid residues in length), which are predominantly composed of very hydrophobic residues. The reagents used for the isolation, fractionation, and characterization of polylysyl-adenosines derived from polylysyl-sRNA include crude preincubated extract of Escherichia coli B harvested 10 minutes after infection with phage T4 at a multiplicity of 5, polyadenylic acid, pancreatic RNase, and L-Lysine-3H. These processes of isolation and characterization are used for polypeptides, which are very basic and not precipitated by phenol, whereas the fractionation procedure is really useful only for polylysine peptides." @default.
- W973404156 created "2016-06-24" @default.
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- W973404156 date "1968-01-01" @default.
- W973404156 modified "2023-09-23" @default.
- W973404156 title "[164] Characterization of the polypeptidyl-sRNA complex in Escherichia coli cell-free systems" @default.
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- W973404156 doi "https://doi.org/10.1016/0076-6879(67)12197-6" @default.
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