Translation and folding of single proteins in real time

Protein biosynthesis is inherently coupled to cotranslational protein folding. Folding of the nascent chain already occurs during synthesis and is mediated by spatial constraints imposed by the ribosomal exit tunnel as well as self-interactions. The polypeptide’s vectorial emergence from the ribosom...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 114; no. 22; pp. E4399 - E4407
Main Authors Wruck, Florian, Katranidis, Alexandros, Nierhaus, Knud H., Büldt, Georg, Hegner, Martin
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 30.05.2017
SeriesPNAS Plus
Subjects
Amino Acid Sequence
Amino acids
Biological Sciences
Biophysical Phenomena
Biosynthesis
Chains
Electrostatic properties
Elongation
Folding
Hydrophobic and Hydrophilic Interactions
Hydrophobicity
Kinetics
Models, Molecular
Optical Tweezers
PNAS Plus
Polypeptides
Proline
Protein Biosynthesis
Protein Folding
Protein Modification, Translational
Protein structure
Proteins
Real time
Ribosomes
Ribosomes - metabolism
Single Molecule Imaging
Stalling
Static Electricity
Synthesis
Tertiary structure
Translation
single molecule
ribosomes
cotranslational protein folding
optical tweezers
protein synthesis
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Summary:Protein biosynthesis is inherently coupled to cotranslational protein folding. Folding of the nascent chain already occurs during synthesis and is mediated by spatial constraints imposed by the ribosomal exit tunnel as well as self-interactions. The polypeptide’s vectorial emergence from the ribosomal tunnel establishes the possible folding pathways leading to its native tertiary structure. How cotranslational protein folding and the rate of synthesis are linked to a protein’s amino acid sequence is still not well defined. Here, we follow synthesis by individual ribosomes using dual-trap optical tweezers and observe simultaneous folding of the nascent polypeptide chain in real time. We show that observed stalling during translation correlates with slowed peptide bond formation at successive proline sequence positions and electrostatic interactions between positively charged amino acids and the ribosomal tunnel. We also determine possible cotranslational folding sites initiated by hydrophobic collapse for an unstructured and two globular proteins while directly measuring initial cotranslational folding forces. Our study elucidates the intricate relationship among a protein’s amino acid sequence, its cotranslational nascent-chain elongation rate, and folding.
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1Present address: Institute for Atomic and Molecular Physics (AMOLF), 1098 XG Amsterdam, The Netherlands.
Edited by George H. Lorimer, University of Maryland, College Park, MD, and approved April 21, 2017 (received for review October 27, 2016)
Author contributions: G.B. conceived the idea of the study; A.K., G.B., and M.H. designed research; F.W. and A.K. performed research; F.W. built the high-resolution optical tweezers and performed the tweezers experiments; A.K. designed and generated all constructs and performed the molecular biology; K.H.N., G.B., and M.H. supervised the study; F.W. and M.H. analyzed data; K.H.N. and G.B. contributed to the writing of the paper; and F.W., A.K., K.H.N., and M.H. wrote the paper.
3Deceased April 7, 2016.
ISSN:0027-8424
1091-6490
1091-6490
DOI:10.1073/pnas.1617873114