title: Site-specific fluorescent labeling of nascent proteins on the translating ribosome.
abstract: As newly synthesized proteins emerge from the ribosome, they interact with a variety of cotranslational cellular machineries that facilitate their proper folding, maturation, and localization. These interactions are essential for proper function of the cell, and the ability to study these events is crucial to understanding cellular protein biogenesis. To this end, we have developed a highly efficient method to generate ribosome-nascent chain complexes (RNCs) site-specifically labeled with a fluorescent dye on the nascent polypeptide. The fluorescent RNC provides real-time, quantitative information on its cotranslational interaction with the signal recognition particle and will be a valuable tool in elucidating the role of the translating ribosome in numerous biochemical pathways.
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, United States.
authors.forename: Ishu
authors.initials: I
authors.lastname: Saraogi
authors.forename: Dawei
authors.initials: D
authors.lastname: Zhang
authors.forename: Sandhya
authors.initials: S
authors.lastname: Chandrasekaran
authors.forename: Shu-ou
authors.initials: SO
authors.lastname: Shan
mesh_terms: D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D008958:Models, Molecular; D015394:Molecular Structure; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000737:chemistry; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins; D012269:Ribosomal Proteins