Functional decoration of elastin-like polypeptides-based nanoparticles with a modular assembly via isopeptide bond...

Area of Science:

• Biomaterials Science
• Protein Engineering
• Nanotechnology

Background:

• Temperature-responsive elastin-like polypeptides (ELPs) undergo phase transitions, making them suitable for nanoparticle construction.
• Existing methods for nanoparticle functionalization can be limited by the thermal processing required for ELP nanoparticle formation.

Purpose of the Study:

• To develop a novel, heat-stable method for decorating ELP-based nanoparticles using the SnoopTag/SnoopCatcher system.
• To demonstrate the successful display and retained activity of functional proteins on these engineered nanoparticles.

Main Methods:

• Constructed protein nanoparticles from ELP-poly(aspartic acid) (ELP-poly(D)) and ELP-poly(D)-SnoopCatcher fusion proteins, leveraging ELP's temperature responsiveness.
• Utilized the covalent isopeptide bond formation of the SnoopTag/SnoopCatcher system for surface functionalization.
• Displayed model proteins, enhanced green fluorescent protein (EGFP) and Renilla luciferace (Rluc), fused to SnoopTag on the nanoparticle surfaces.

Main Results:

• The SnoopTag/SnoopCatcher system enabled nanoparticle decoration without compromising particle formation due to heat.
• EGFP retained 48.7% of its activity, and Rluc retained nearly full activity after surface display on the nanoparticles.
• The method proved effective for attaching proteins with varying thermostability.

Conclusions:

• The ELP-based nanoparticles with the SnoopTag/SnoopCatcher system provide a versatile platform for heat-stable protein immobilization.
• This approach facilitates the decoration of nanoparticles with diverse functional proteins via robust isopeptide bond formation.
• The developed method expands the utility of ELP-based nanoparticles in various applications requiring stable protein conjugation.