Slowing Bacterial Translation Speed Enhances Eukaryotic Protein Folding Efficiency

Efraín Siller, Diane C. DeZwaan, John F. Anderson, Brian C. Freeman, José M. Barral

    Research output: Contribution to journalArticle

    101 Scopus citations

    Abstract

    The mechanisms for de novo protein folding differ significantly between bacteria and eukaryotes, as evidenced by the often observed poor yields of native eukaryotic proteins upon recombinant production in bacterial systems. Polypeptide synthesis rates are faster in bacteria than in eukaryotes, but the effects of general variations in translation rates on protein folding efficiency have remained largely unexplored. By employing Escherichia coli cells with mutant ribosomes whose translation speed can be modulated, we show here that reducing polypeptide elongation rates leads to enhanced folding of diverse proteins of eukaryotic origin. These results suggest that in eukaryotes, protein folding necessitates slow translation rates. In contrast, folding in bacteria appears to be uncoupled from protein synthesis, explaining our findings that a generalized reduction in translation speed does not adversely impact the folding of the endogenous bacterial proteome. Utilization of this strategy has allowed the production of a native eukaryotic multidomain protein that has been previously unattainable in bacterial systems and may constitute a general alternative to the production of aggregation-prone recombinant proteins.

    Original languageEnglish (US)
    Pages (from-to)1310-1318
    Number of pages9
    JournalJournal of Molecular Biology
    Volume396
    Issue number5
    DOIs
    StatePublished - 2010

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    Keywords

    • Bacteria
    • Eukaryotes
    • Protein folding
    • Protein synthesis
    • Recombinant proteins

    ASJC Scopus subject areas

    • Molecular Biology

    Cite this

    Siller, E., DeZwaan, D. C., Anderson, J. F., Freeman, B. C., & Barral, J. M. (2010). Slowing Bacterial Translation Speed Enhances Eukaryotic Protein Folding Efficiency. Journal of Molecular Biology, 396(5), 1310-1318. https://doi.org/10.1016/j.jmb.2009.12.042