A general approach to protein folding using thermostable exoshells

Area of Science:

• Biochemistry
• Nanotechnology
• Protein Science

Background:

• In vitro protein folding is challenging, often leading to aggregation, low yields, and reduced specific activity.
• Existing methods struggle with diverse protein types, including those with complex structures or multiple subunits.

Purpose of the Study:

• To investigate the use of nanoscale exoshells (tES) as complementary nanoenvironments for in vitro protein folding.
• To assess the impact of tES on the yield, activity, and solubility of a diverse range of protein substrates.

Main Methods:

• Encapsulation of 12 diverse protein substrates within tES.
• Evaluation of protein folding, yield, and specific activity after release from tES.
• Analysis of factors influencing functional folding, such as charge complementation.

Main Results:

• Protein encapsulation within tES increased soluble yield (3-fold to >100-fold), functional yield (2-fold to >100-fold), and specific activity (3-fold to >100-fold) for all tested proteins.
• Average soluble yield was 6.5 mg/100 mg of tES.
• Charge complementation between tES internal cavity and protein substrate was identified as the primary determinant of functional folding.

Conclusions:

• Nanoscale exoshells provide an effective solution for improving in vitro protein folding.
• Electrostatic interactions at the nanoscale play a crucial role in successful protein folding within tES.
• This technology offers a significant advancement for producing functional proteins in vitro.