title: Peptidyl transferase center activity observed in single ribosomes.
abstract: We demonstrate the functional activity of single ribosomal complexes, opening the way for detailed studies of the trajectories of protein synthesis. Our approach employs a single-molecule detection system, capable of picoseconds to minutes resolution, to observe a growing peptide labeled at its N terminus with the fluorophore tetramethylrhodamine (TMR). Single complexes of mRNA-programmed ribosomes with TMR-Met-tRNAMetf or TMR-Met-Phe-tRNAPhe are immobilized on mica and observed by fluorescence. Immobilized ribosome.mRNA.TMR-Met-tRNAMetf complexes form peptide bonds with puromycin. Single-molecule detection reveals dynamics on the scale of seconds at the ribosomal peptidyl transferase center.
authors.affiliation: Department of Chemistry, University of Pennsylvania, Philadelphia, PA, 19104, USA.
authors.forename: A
authors.initials: A
authors.lastname: Sytnik
authors.forename: S
authors.initials: S
authors.lastname: Vladimirov
authors.forename: Y
authors.initials: Y
authors.lastname: Jia
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.forename: B S
authors.initials: BS
authors.lastname: Cooperman
authors.forename: R M
authors.initials: RM
authors.lastname: Hochstrasser
mesh_terms: D005456:Fluorescent Dyes; D010458:Peptidyl Transferases / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000378:metabolism; D012235:Rhodamines; D012270:Ribosomes / Q000201:enzymology*
chemical_list: D005456:Fluorescent Dyes; C072113:Met-tRNA(f)(Met); D012358:RNA, Transfer, Met; D012360:RNA, Transfer, Phe; D012235:Rhodamines; C005358:tetramethylrhodamine; D010458:Peptidyl Transferases