Tandem-repeat proteins introduce tuneable properties to engineered biomolecular condensates

Area of Science:

• Cell biology
• Biochemistry
• Synthetic biology

Background:

• Biomolecular condensates are crucial for cellular functions.
• Understanding condensate structure-function relationships requires controllable synthetic systems.

Purpose of the Study:

• To design and characterize a tuneable synthetic biomolecular condensate platform.
• To investigate the link between CTPR structural attributes and condensate properties.
• To enable specific target-protein recruitment into synthetic condensates.

Main Methods:

• Fusion of modular consensus-designed tetratricopeptide repeat (CTPR) proteins with intrinsically-disordered domains.
• Experimental characterization of condensate properties.
• In silico modeling to analyze structure-propensity relationships.
• Incorporation of short binding motifs for target recruitment.

Main Results:

• CTPR structural attributes systematically and predictably influence condensate propensity.
• Experimental and in silico data confirmed the trends between CTPR structure and condensate behavior.
• Synthetic condensates demonstrated specific recruitment of target proteins via incorporated binding motifs.

Conclusions:

• The developed synthetic condensate platform offers tuneable control over condensate properties.
• This system can be rationally designed to impart new functions, such as specific protein recruitment.
• The platform serves as a versatile tool for studying biomolecular condensates and their biological roles.