Modular assembly of proteins on nanoparticles

Abstract

Generally, the high diversity of protein properties necessitates the development of unique nanoparticle bio-conjugation methods, optimized for each different protein. Here we describe a universal bio-conjugation approach which makes use of a new recombinant fusion protein combining two distinct domains. The N-terminal part is Glutathione S-Transferase (GST) from Schistosoma japonicum, for which we identify and characterize the remarkable ability to bind gold nanoparticles (GNPs) by forming gold–sulfur bonds (Au–S). The C-terminal part of this multi-domain construct is the SpyCatcher from Streptococcus pyogenes, which provides the ability to capture recombinant proteins encoding a SpyTag. Here we show that SpyCatcher can be immobilized covalently on GNPs through GST without the loss of its full functionality. We then show that GST-SpyCatcher activated particles are able to covalently bind a SpyTag modified protein by simple mixing, through the spontaneous formation of an unusual isopeptide bond.

Post‐translational Assembly of Protein Parts into Complex Devices by Using SpyTag/SpyCatcher Protein Ligase

“…Conjugating proteins to nanoparticles typically necessitates optimization for each protein. Ma et al . have described a universal strategy using SpyTag‐modified proteins and SpyCatcher immobilized through glutathione S‐transferase (GST) to gold nanoparticles (AuNPs), thus creating protein‐decorated AuNPs, which serve a variety of functions, from biosensors to nanomaterials and nanomedicine.…”

A Plug and Play Approach for the Decoration of Nanoparticles With Recombinant Proteins

“…The tag would allow the end user to 'plug' the protein onto any nanoparticle for which an 'adaptor' exists. A molecular equivalent of this adaptor has been recently described [13] for the use with gold nanoparticles. Gold nanoparticles were chosen as a proof-of-concept and for their general relevance in nanomedicine [15].…”

“…This is a common affinity tag used in protein biochemistry for its ability to bind glutathione-modified resins. It was discovered that GST also adsorbs onto citrate-capped gold nanoparticles with remarkable affinity and the mechanism of binding was described [13]. This involves the formation of covalent bonds between gold and sulfur atoms at cysteine residues (Au-S bonds), which make the binding of GST to gold permanent.…”

“…As the excessive diversification of methods and chemistries in nanocarriers' synthesis could potentially lead to regulatory complications, it is important to study modular strategies to simplify and reduce the number of molecular building blocks required. Such approach was recently reported in the context of modular assembly of proteins on gold nanoparticles [13] and it is detailed below.…”

Novel fluorescein polymer-based nanoparticles: facile and controllable one-pot synthesis, assembly, and immobilization of biomolecules for application in a highly sensitive biosensor

“…1,10,14 In addition, to enhance the diagnostic specicity of nanoparticle-based assays, binding of the target biomolecule to the NPs is essential. 21,22 For the successful detection of the target analyte, this binding should result in a measurable signal that can be quantied. The presence of reactive functional groups along the polymer backbone therefore allows specic labeling with other molecules to be carried out, in addition to modication of the particle surface for specic applications.…”