title: Mutational analysis of protein folding inside the ribosome exit tunnel.
abstract: Recent work has demonstrated that cotranslational folding of proteins or protein domains in, or in the immediate vicinity of, the ribosome exit tunnel generates a pulling force on the nascent polypeptide chain that can be detected using a so-called translational arrest peptide (AP) engineered into the nascent chain as a force sensor. Here, we show that AP-based force measurements combined with systematic Ala and Trp scans of a zinc-finger domain that folds in the exit tunnel can be used to identify the residues that are critical for intraribosomal folding. Our results suggest a general approach to characterize the folded state(s) that may form as a protein domain moves progressively down the ribosome exit tunnel.
authors.affiliation: Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, Sweden.
authors.forename: José Arcadio
authors.initials: JA
authors.lastname: Farías-Rico
authors.affiliation: Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, Sweden.
authors.forename: Sara Kathrin
authors.initials: SK
authors.lastname: Goetz
authors.affiliation: Gene Center and Center for Integrated Protein Science Munich, CiPS-M, University of Munich, Germany.
authors.forename: Jacopo
authors.initials: J
authors.lastname: Marino
authors.affiliation: Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, Sweden.|Science for Life Laboratory, Stockholm University, Solna, Sweden.
authors.forename: Gunnar
authors.initials: G
authors.lastname: von Heijne
mesh_terms: D000409:Alanine / Q000378:metabolism; D000595:Amino Acid Sequence; D001696:Biomechanical Phenomena; D002942:Circular Dichroism; D004252:DNA Mutational Analysis; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D010455:Peptides / Q000378:metabolism; D000072417:Protein Domains; D017510:Protein Folding; D012270:Ribosomes / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
chemical_list: D010455:Peptides; D014364:Tryptophan; D015032:Zinc; D000409:Alanine
keywords: arrest peptide; cotranslational protein folding; ribosome