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Farias-Rico, Jose Arcadio; Götz, Sara Kathrin; Marino, Jacopo; Heijne, Gunnar von (2017): Mutational analysis of protein folding inside the ribosome exit tunnel. In: Febs Letters, Vol. 591, No. 1: pp. 155-163
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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.