Slowing bacterial translation speed enhances eukaryotic protein folding efficiency

Area of Science:

• Molecular Biology
• Biochemistry
• Protein Engineering

Background:

• Eukaryotic and bacterial protein folding mechanisms differ, leading to low yields of recombinant eukaryotic proteins in bacteria.
• Faster bacterial polypeptide synthesis rates may hinder eukaryotic protein folding efficiency.
• The impact of translation speed on protein folding efficiency is not well understood.

Purpose of the Study:

• To investigate the effect of modulating translation rates on eukaryotic protein folding in bacteria.
• To determine if slower translation speeds can improve the production of soluble eukaryotic proteins.
• To explore strategies for overcoming challenges in recombinant protein production.

Main Methods:

• Utilized Escherichia coli with mutant ribosomes to control polypeptide elongation rates.
• Compared folding efficiency of diverse eukaryotic proteins at varying translation speeds.
• Assessed the impact of reduced translation speed on endogenous bacterial proteome folding.

Main Results:

• Decreasing polypeptide elongation rates significantly enhanced the folding of eukaryotic proteins.
• Slower translation is necessary for efficient eukaryotic protein folding.
• Bacterial protein folding is largely independent of translation speed.
• Successfully produced a previously unattainable native eukaryotic multidomain protein.

Conclusions:

• Bacterial systems can be engineered for improved eukaryotic protein folding by slowing translation.
• This strategy offers a novel approach for producing aggregation-prone recombinant proteins.
• Understanding the interplay between translation speed and protein folding is key for biotechnological applications.