Abstract: Disclosed herein are methods and kits useful in the detection of protein folding and in the identification of compounds that promote proper protein folding. In one example approach, fluorophores and a protein tag are incorporated into a nascent polypeptide within a ribosome-nascent-chain complex during cell free translation and the resulting labeled ribosome-nascent-chain complex is conjugated to a solid surface via the tag. Fluorescence imaging via FRET is then preformed to assess the folding state of the ribosome-nascent-chain complex under a variety of conditions.

Abstract: Disclosed herein are methods and kits useful in the detection of protein folding and in the identification of compounds that promote proper protein folding. In one example approach, fluorophores and a protein tag are incorporated into a nascent polypeptide within a ribosome-nascent-chain complex during cell free translation and the resulting labeled ribosome-nascent-chain complex is conjugated to a solid surface via the tag. Fluorescence imaging via FRET is then preformed to assess the folding state of the ribosome-nascent-chain complex under a variety of conditions.

Abstract: Disclosed herein are compositions and methods for incorporating non-native amino acids into a single polypeptide. In one example, an isolated modified suppressor tRNA is disclosed that includes the following: a modified tRNAcys, wherein the tRNAcys has been modified so that an anticodon of the tRNA is complementary to a stop codon; a cysteine amino acid residue is covalently linked to the modified tRNAcys by aminoacylation generating a chemically reactive sulfhydryl side chain; and a detectable label is covalently linked to the sulfhydryl side chain. Methods for incorporating non-native amino acids into a single polypeptide utilizing the disclosed isolated modified suppressor tRNAs are also disclosed.