Slowing bacterial translation speed enhances eukaryotic protein folding efficiency.
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.
Siller
Efraín
E
Department of Neuroscience and Cell Biology, The University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-0620, USA.
DeZwaan
Diane C
DC
Anderson
John F
JF
Freeman
Brian C
BC
Barral
José M
JM
D000818:Animals; D020240:Apraxia, Ideomotor; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D066298:In Vitro Techniques; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D009154:Mutation; D010441:Peptide Chain Elongation, Translational; D014176:Protein Biosynthesis; D017510:Protein Folding; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012269:Ribosomal Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics
D017931:DNA Primers; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C506766:RpsL protein, E coli; D049452:Green Fluorescent Proteins; D049409:Luciferases, Firefly