Rational design of proteins that exchange on functional timescales

Area of Science:

• Protein engineering
• Computational biology
• Biophysics

Background:

• Proteins are dynamic molecules essential for biological functions, existing in multiple conformational states.
• Current protein design methods struggle to replicate natural efficiencies due to limitations in controlling conformational exchange on functional timescales.

Purpose of the Study:

• To develop a computational method for engineering protein dynamics, enabling control over conformational exchange.
• To design proteins that can spontaneously access multiple conformational states on functionally relevant timescales.

Main Methods:

• Developed a computational method termed meta-multistate design.
• Applied this method to engineer Streptococcal protein G domain β1.
• Introduced two novel conformations into the protein's global fold.

Main Results:

• Successfully designed proteins, named DANCERs (dynamic and native conformational exchangers), that are stably folded.
• Demonstrated spontaneous conformational exchange between the designed states on the millisecond timescale.
• Validated the computational method's ability to engineer protein dynamics.

Conclusions:

• Meta-multistate design is a broadly applicable method for engineering protein dynamics.
• The ability to control conformational exchange on functional timescales opens new avenues for protein design and applications.
• DANCER proteins represent a significant advancement in creating functional proteins with tailored dynamic properties.