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ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly

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A Self-Assembling Two-Dimensional Protein Array is a Versatile Platform for the Assembly of Multicomponent

Alexander Thomas¹, James F Matthaei¹, François Baneyx¹

¹Department of Chemical Engineering, University of Washington, Box 351750, Seattle, WA 98195, USA.

Biotechnology Journal

|September 1, 2018

Summary

Researchers modified a self-assembling protein (TTM) to create versatile two-dimensional (2D) scaffolds. These protein arrays can be functionalized for diverse nanoscale applications in catalysis, sensing, and biomedicine.

Keywords:

2D arrays 2D lattices nanofabrication self-assembly solid binding proteins solid-binding peptides

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Area of Science:

Biomaterials science
Nanotechnology
Protein engineering

Background:

Two-dimensional (2D) arrays are crucial for nanoscale component assembly in catalysis, sensing, and biomedical fields.
Computational redesign previously yielded the TTM protein, which self-assembles into micrometer-scale nanostructured lattices.
The accessibility of TTM monomer termini was not previously characterized.

Purpose of the Study:

To investigate the functionalization potential of the TTM protein scaffold.
To demonstrate the use of modified TTM for creating hierarchical nanostructures.
To explore applications in nanoparticle conjugation and controlled biomolecule immobilization.

Main Methods:

Computational protein redesign of TTM for calcium-induced self-assembly.
Modification of TTM N- and C-termini with hexahistidine, gold-binding peptide, and biotinylation tags.
Assembly of modified TTM onto surfaces and co-polymerization with wild-type TTM.

Main Results:

The N- and C-termini of TTM monomers were confirmed as solvent-accessible.
Functionalized TTM successfully decorated beads with protein islands, conjugated gold nanoparticles, and controlled avidin immobilization.
Co-polymerization enabled tunable immobilization densities of avidin onto 2D lattices.

Conclusions:

The TTM protein serves as a versatile 2D scaffold for fabricating hierarchical structures.
Tailored functionalization of TTM opens possibilities for advanced nanomaterial design.
This work highlights TTM's potential in catalysis, sensing, and biomedical engineering.