Production of Multicomponent Protein Templates for the Positioning and Stabilization of Enzymes

Part of the Methods in Molecular Biology book series (MIMB, volume 2073)


Harnessing the ability of proteins to self-assemble into complex structures has enabled the creation of templates for applications in nanotechnology. Protein templates can be used to position functional molecules in regular patterns with nanometer precision over large surface areas. A difficult but successful approach to building customizable protein templates involves designing novel protein-protein interfaces to join protein building blocks into ordered arrangements. This approach was illustrated recently by engineering the protein interfaces of a molecular chaperone to produce filamentous templates composed of repeating subunits. In this chapter, we describe how these multicomponent protein templates can be produced recombinantly, assembled into filaments, and used as material templates. The templates enable the positioning and alignment of functional molecules at varying distances along the length of the filament, which can be demonstrated using a Förster resonance energy transfer (FRET) assay. In addition, we describe a method to quantify the chaperone ability of these filaments to stabilize and protect other proteins from thermal-induced aggregation—a useful property for bionanotechnology applications that involve molecular scaffolds for positioning and stabilizing enzymes.

Key words

Filament Self-assembly Protein interface Coiled-coil Scaffold Template Chaperone Biomaterials 



This work was supported by the Air Force Office of Scientific Research (FA9550-14-1-0026).


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Chemical and Biomolecular EngineeringUniversity of CaliforniaBerkeleyUSA
  2. 2.School of Biotechnology and Biomolecular SciencesThe University of New South WalesSydneyAustralia

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