In vitro protein synthesis directed by R17 viral ribonucleic acid. III. A new assay method for cell-free protein synthesis based on a phenylthiohydantoin procedure

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Abstract
A new method of assaying cell-free polypeptide synthesis was sought, in order to make it possible to measure peptide synthesis regardless of the solubility of products in aqueous solvent. By direct application of Edman's degradation procedure, the phenylthiohydantoins of free unreacted amino acids, as well as of amino terminal residues of peptides synthesized in a cell-free system, were extracted, and the quantity of amino acids incorporated into peptide was measured. With this method (liquid-phase method), it was discovered that in our cell-free system, lysine and phenylalanine were apparently polymerized at the same rate in the presence of the corresponding mRNA. There was also an appreciable amount of amino acid polymerization directed by R17 viral RNA whose products are mostly soluble in acid. The liquid-phase method was successfully modified so that all assay procedures could take place on filter paper. Thus it became possible to process a large number of samples simultaneously.

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